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mirVana™ PARIS™ RNA and Native Protein Purification Kit (Invitrogen™)
The mirVana™ PARIS™ Kit is for the purification of both native protein and RNA (including small RNA) from the same sample. Each kit contains sufficient reagents and consumables for either 40 isolations of protein and total RNA (including small RNAs), or 20 isolations of protein and separate large and small RNA fractions. Features of this kit:
• Isolate small RNA-containing total RNA and native protein from the same sample
• Start with 100–107 cultured cells or up to 100 mg of many mammalian tissues
• Enrich for small RNA (<200 nt) to increase sensitivity in downstream analyses
• Follow a simple, 30-minute procedure
• Use to correlate mRNA, miRNA or siRNA, and protein levels
How the kit works
The mirVana™ PARIS™ Kit procedure begins with homogenization of samples with a special cell disruption buffer that includes nonionic detergent. Protein remains intact, so a portion of the lysate can be used directly for common protein analysis applications. The remainder of the lysate is used for RNA isolation, using a procedure that combines the advantages of organic and solid-phase extraction, while avoiding the disadvantages of both.
Accessory products
For tissues that have very high levels of ribonucleases, such as pancreas or spleen, the mirVana™ miRNA Isolation Kit is recommended, although it is not designed to recover protein and small RNA from the same sample.
• Isolate small RNA-containing total RNA and native protein from the same sample
• Start with 100–107 cultured cells or up to 100 mg of many mammalian tissues
• Enrich for small RNA (<200 nt) to increase sensitivity in downstream analyses
• Follow a simple, 30-minute procedure
• Use to correlate mRNA, miRNA or siRNA, and protein levels
How the kit works
The mirVana™ PARIS™ Kit procedure begins with homogenization of samples with a special cell disruption buffer that includes nonionic detergent. Protein remains intact, so a portion of the lysate can be used directly for common protein analysis applications. The remainder of the lysate is used for RNA isolation, using a procedure that combines the advantages of organic and solid-phase extraction, while avoiding the disadvantages of both.
Accessory products
For tissues that have very high levels of ribonucleases, such as pancreas or spleen, the mirVana™ miRNA Isolation Kit is recommended, although it is not designed to recover protein and small RNA from the same sample.
B-PER™ Bacterial Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific B-PER Bacterial Protein Extraction Reagent is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
Features of the B-PER Bacterial Protein Extraction Reagent:
• Detergent in Tris buffer; no enzyme components
• Bacterial lysis
• Purification of affinity tagged proteins
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Complete Bacterial Protein Extraction Reagent
B-PER™ Reagent (in Phosphate Buffer)
B-PER™ II Bacterial Protein Extraction Reagent (2X)
Features of the B-PER Bacterial Protein Extraction Reagent:
• Detergent in Tris buffer; no enzyme components
• Bacterial lysis
• Purification of affinity tagged proteins
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Complete Bacterial Protein Extraction Reagent
B-PER™ Reagent (in Phosphate Buffer)
B-PER™ II Bacterial Protein Extraction Reagent (2X)
B-PER™ II Bacterial Protein Extraction Reagent (2X) (Thermo Scientific™)
Thermo Scientific B-PER II Bacterial Protein Extraction Reagent (2X) is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
Features of the B-PER Bacterial Protein Extraction Reagent:
• 2X B-PER (detergent in Tris buffer; no enzyme components)
• Bacterial lysis
• Purification of proteins having low expression levels
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent
B-PER™ Reagent (in Phosphate Buffer)
Features of the B-PER Bacterial Protein Extraction Reagent:
• 2X B-PER (detergent in Tris buffer; no enzyme components)
• Bacterial lysis
• Purification of proteins having low expression levels
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent
B-PER™ Reagent (in Phosphate Buffer)
P-PER™ Plant Protein Extraction Reagent (Thermo Scientific™)
The Thermo Scientific P-PER Plant Protein Extraction Kit effectively extracts protein from all kinds of dry and fresh plant tissue without liquid nitrogen.
Features of the P-PER Plant Protein Extraction Kit:
• Versatile—works with multiple plant organs (leaf, stem, root, seed and flowers); multiple plant species (Arabidopsis, tobacco, maize, soybeans, peas, spinach and other plant tissues); and fresh, frozen and dehydrated plant tissues
• Convenient—requires no liquid nitrogen/freeze-grinding, Dounce homogenization, blade-shearing or glass-bead agitation for cell disruption; however, the P-PER Kit is compatible with these alternative mechanical aids
• Compatible—downstream applications include 1-D and 2-D gel electrophoresis, Western blotting, activity assays, and protein affinity purifications
• Quantifiable—P-PER Kit extracts can be quantified using the BCA Protein Assay Kit, Reducing Agent Compatible (Part No. 23250)
• Ready-to-use—protein extract does not require filtration through cheesecloth or Miracloth, unlike homebrews
• Fast—perform plant cell lysis and protein extraction in 10 minutes
• Recovers active protein—assays show extracted proteins are functional
The P-PER Kit includes an organic lysing reagent and two aqueous reagents that effectively lyse plant cells and solubilize protein without harsh mechanical lysis aids, such as a mortar and pestle. Plant protein extracts are prepared in just 10 minutes, and protein yields exceed conventional extraction methods (e.g., freeze/grinding in liquid nitrogen) and other commercially available plant protein extraction reagents. The resulting protein extracts are compatible with a variety of downstream applications including the Thermo Scientific BCA Protein Assay Kit - Reducing Agent Compatible (Part No. 23250), 1D and 2D gel electrophoresis, immunoprecipitation, Western blotting, activity assays and protein purification.
Features of the P-PER Plant Protein Extraction Kit:
• Versatile—works with multiple plant organs (leaf, stem, root, seed and flowers); multiple plant species (Arabidopsis, tobacco, maize, soybeans, peas, spinach and other plant tissues); and fresh, frozen and dehydrated plant tissues
• Convenient—requires no liquid nitrogen/freeze-grinding, Dounce homogenization, blade-shearing or glass-bead agitation for cell disruption; however, the P-PER Kit is compatible with these alternative mechanical aids
• Compatible—downstream applications include 1-D and 2-D gel electrophoresis, Western blotting, activity assays, and protein affinity purifications
• Quantifiable—P-PER Kit extracts can be quantified using the BCA Protein Assay Kit, Reducing Agent Compatible (Part No. 23250)
• Ready-to-use—protein extract does not require filtration through cheesecloth or Miracloth, unlike homebrews
• Fast—perform plant cell lysis and protein extraction in 10 minutes
• Recovers active protein—assays show extracted proteins are functional
The P-PER Kit includes an organic lysing reagent and two aqueous reagents that effectively lyse plant cells and solubilize protein without harsh mechanical lysis aids, such as a mortar and pestle. Plant protein extracts are prepared in just 10 minutes, and protein yields exceed conventional extraction methods (e.g., freeze/grinding in liquid nitrogen) and other commercially available plant protein extraction reagents. The resulting protein extracts are compatible with a variety of downstream applications including the Thermo Scientific BCA Protein Assay Kit - Reducing Agent Compatible (Part No. 23250), 1D and 2D gel electrophoresis, immunoprecipitation, Western blotting, activity assays and protein purification.
B-PER™ with Enzymes Bacterial Protein Extraction Kit (Thermo Scientific™)
Thermo Scientific B-PER with Enzymes Bacterial Protein Extraction Kit is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
Features of the B-PER Bacterial Protein Extraction Reagent:
• Tris buffered formulation with lysozyme and DNase enzymes
• Improved cell membrane solubilization and DNA digestion
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent
Features of the B-PER Bacterial Protein Extraction Reagent:
• Tris buffered formulation with lysozyme and DNase enzymes
• Improved cell membrane solubilization and DNA digestion
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent
Mem-PER™ Plus Membrane Protein Extraction Kit (Thermo Scientific™)
The Thermo Scientific Mem-PER Plus Membrane Protein Extraction Kit enables small-scale solubilization and enrichment of integral membrane proteins and membrane-associated proteins in a simple reagent-based procedure.
Features of the Mem-PER Plus Kit:
• Extraction and isolation—produces minimal cross-contamination (typically less than 10%) of cytosolic protein into the membrane protein fraction
• Cells or tissues—effective for extraction from cultured mammalian cells and mammalian tissues
• Downstream compatibility—analyze membrane protein extracts by SDS-PAGE, western blotting, immunoprecipitation and protein assays
• Fast and simple—isolation of membrane proteins in approximately one hour
• No special equipment required—only a benchtop microcentrifuge, tubes, homogenizers, and pipettors are needed
The Mem-PER Plus Kit effectively isolates membrane proteins from cultured cells and tissues using a simple benchtop microcentrifuge procedure. Integral membrane proteins having one or two membrane-spanning domains are effectively solubilized and isolated from cytosolic proteins. The kit makes possible the extraction and selective enrichment of integral and attached membrane proteins from cultured mammalian cells, as well as from hard and soft mammalian tissues. The Mem-PER Plus Kit has many advantages compared to our original Mem-PER Kit. The protocol is simpler, and the fractions are directly compatible with many downstream applications such as SDS-PAGE, Western Blotting, BCA, immunoprecipitation, and amine-reactive protein labeling techniques.
Includes:
Kit contains a cell wash solution, permeabilization buffer, and solubilization buffer.
Traditional methods for isolation of membrane protein are tedious and time-consuming, and they require gradient separation and expensive ultracentrifugation equipment. The Mem-PER Plus Membrane Protein Extraction Kit is for the enrichment of integral membrane proteins and membrane associated proteins from cultured mammalian cells or tissue using a mild detergent-based selective extraction protocol. The use of selective detergent extraction eliminates the hassle of phase separation based on hydrophobicity, allowing better reproducibility and higher throughput.
The cells are first permeabilized with a mild detergent, allowing the release of soluble cytosolic proteins, after which a second detergent solubilizes membrane proteins. Extraction efficiencies and yields will vary depending on cell type as well as the number of times the integral membrane protein spans the lipid bilayer. Membrane proteins with one or two transmembrane domains are typically extracted with an efficiency of up to 90%. Cross-contamination of cytosolic proteins into the membrane fraction is usually less than 10%.
More Product Data
• Efficient mammalian membrane protein extraction
Related Products
Subcellular Protein Fractionation Kit for Cultured Cells
Subcellular Protein Fractionation Kit for Tissues
Features of the Mem-PER Plus Kit:
• Extraction and isolation—produces minimal cross-contamination (typically less than 10%) of cytosolic protein into the membrane protein fraction
• Cells or tissues—effective for extraction from cultured mammalian cells and mammalian tissues
• Downstream compatibility—analyze membrane protein extracts by SDS-PAGE, western blotting, immunoprecipitation and protein assays
• Fast and simple—isolation of membrane proteins in approximately one hour
• No special equipment required—only a benchtop microcentrifuge, tubes, homogenizers, and pipettors are needed
The Mem-PER Plus Kit effectively isolates membrane proteins from cultured cells and tissues using a simple benchtop microcentrifuge procedure. Integral membrane proteins having one or two membrane-spanning domains are effectively solubilized and isolated from cytosolic proteins. The kit makes possible the extraction and selective enrichment of integral and attached membrane proteins from cultured mammalian cells, as well as from hard and soft mammalian tissues. The Mem-PER Plus Kit has many advantages compared to our original Mem-PER Kit. The protocol is simpler, and the fractions are directly compatible with many downstream applications such as SDS-PAGE, Western Blotting, BCA, immunoprecipitation, and amine-reactive protein labeling techniques.
Includes:
Kit contains a cell wash solution, permeabilization buffer, and solubilization buffer.
Traditional methods for isolation of membrane protein are tedious and time-consuming, and they require gradient separation and expensive ultracentrifugation equipment. The Mem-PER Plus Membrane Protein Extraction Kit is for the enrichment of integral membrane proteins and membrane associated proteins from cultured mammalian cells or tissue using a mild detergent-based selective extraction protocol. The use of selective detergent extraction eliminates the hassle of phase separation based on hydrophobicity, allowing better reproducibility and higher throughput.
The cells are first permeabilized with a mild detergent, allowing the release of soluble cytosolic proteins, after which a second detergent solubilizes membrane proteins. Extraction efficiencies and yields will vary depending on cell type as well as the number of times the integral membrane protein spans the lipid bilayer. Membrane proteins with one or two transmembrane domains are typically extracted with an efficiency of up to 90%. Cross-contamination of cytosolic proteins into the membrane fraction is usually less than 10%.
More Product Data
• Efficient mammalian membrane protein extraction
Related Products
Subcellular Protein Fractionation Kit for Cultured Cells
Subcellular Protein Fractionation Kit for Tissues
M-PER™ Mammalian Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific M-PER Mammalian Protein Extraction Reagent is designed to provide highly efficient total soluble protein extraction from cultured mammalian cells.
Features of M-PER Mammalian Protein Extraction Reagent:
• Gentle—mild detergent lysis, yielding extracts that are immediately compatible with coomassie (Bradford) and BCA protein assays or SDS-PAGE
• Compatible—extracts soluble proteins in nondenatured state, enabling direct use in immunoprecipitation and other affinity purification procedures
• Amine-free and dialyzable—formulation ensures compatibility with subsequent assay systems
• Convenient—lyse adherent cells directly in plate or after scraping and washing in suspension
• Non-denaturing—maintain activity of luciferase, beta-galactosidase, CAT and other reporter genes as well or better than other suppliers' products and freeze/thaw methods
Product Details
The complete cell lysis reagent contains a mild, nondenaturing detergent that dissolves cell membranes to extract and solubilize total protein from most cellular compartments. Extraction is accomplished in only 5 minutes and requires little or no additional mechanical disruption. M-PER reagent is formulated for minimal interference with downstream biological applications. The reagent has been validated for use with several cell lines, including primary, suspension and adherent cell types; the resulting cell lysates are compatible with many downstream assays including immunoassays, enzyme assays and a variety of common reporter assays.
Cellular protein extraction—cell lysis to release the proteins of interest—is a key first step in many proteomics analysis procedures. M-PER reagent is designed to efficiently extract soluble protein from a variety of cell types, including primary cells and cells grown in suspension or adherent culture conditions. Performance of M-PER reagent was evaluated for yields of both total protein and specific target proteins from the various cellular compartments. Additionally, because M-PER reagent is formulated with dialyzable components that are devoid of primary amines, it is more compatible with certain downstream applications, such as luciferase, beta-galactosidase, and CAT assays, than other protein extraction methods.
Related Products
T-PER™ Tissue Protein Extraction Reagent
Features of M-PER Mammalian Protein Extraction Reagent:
• Gentle—mild detergent lysis, yielding extracts that are immediately compatible with coomassie (Bradford) and BCA protein assays or SDS-PAGE
• Compatible—extracts soluble proteins in nondenatured state, enabling direct use in immunoprecipitation and other affinity purification procedures
• Amine-free and dialyzable—formulation ensures compatibility with subsequent assay systems
• Convenient—lyse adherent cells directly in plate or after scraping and washing in suspension
• Non-denaturing—maintain activity of luciferase, beta-galactosidase, CAT and other reporter genes as well or better than other suppliers' products and freeze/thaw methods
Product Details
The complete cell lysis reagent contains a mild, nondenaturing detergent that dissolves cell membranes to extract and solubilize total protein from most cellular compartments. Extraction is accomplished in only 5 minutes and requires little or no additional mechanical disruption. M-PER reagent is formulated for minimal interference with downstream biological applications. The reagent has been validated for use with several cell lines, including primary, suspension and adherent cell types; the resulting cell lysates are compatible with many downstream assays including immunoassays, enzyme assays and a variety of common reporter assays.
Cellular protein extraction—cell lysis to release the proteins of interest—is a key first step in many proteomics analysis procedures. M-PER reagent is designed to efficiently extract soluble protein from a variety of cell types, including primary cells and cells grown in suspension or adherent culture conditions. Performance of M-PER reagent was evaluated for yields of both total protein and specific target proteins from the various cellular compartments. Additionally, because M-PER reagent is formulated with dialyzable components that are devoid of primary amines, it is more compatible with certain downstream applications, such as luciferase, beta-galactosidase, and CAT assays, than other protein extraction methods.
Related Products
T-PER™ Tissue Protein Extraction Reagent
T-PER™ Tissue Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific T-PER Tissue Protein Extraction Reagent enables mild extraction of total protein from tissue samples.
Features of T-PER Tissue Protein Extraction Reagent:
• Simple procedure—just homogenize tissue sample in 1:20 (w/v) of tissue to T-PER Reagent, then centrifuge to pellet cell or tissue debris
• Mild and convenient—detergent is dialyzable for quick and easy removal; can be used with additional components (e.g., protease inhibitors, salts, reducing agents, chelating agents, etc.)
• Flexible—lysate may be used for reporter assays, protein kinase assays, immunoassays and/or protein purifications
• Compatible—the lysate is compatible with standard protein assays such as Pierce Coomassie Plus (Bradford) Protein Assay
Product Details
This tissue cell lysis reagent utilizes a proprietary detergent in 25mM bicine, 150mM sodium chloride (pH 7.6) to maximize the efficiency of protein solubilization from mammalian tissue samples by homogenization. The simple composition is compatible with additives such as protease inhibitors, salts, reducing agents, chelating agents, providing versatility for many different sample types and lysis applications. Cell lysates prepared with T-PER Reagent are directly compatible with reporter assays (e.g., luciferase, beta-galactosidase, chloramphenicol acetyl transferase), protein kinase assays (e.g., PKA, PKC, tyrosine kinase), immunoassays (e.g., Western blots, ELISAs, RIAs) and/or protein purification procedures.
Homogeneous and efficient tissue protein extraction from different samples is challenging due to varying tissue composition and extraction methods. Fibrous and collagenous tissues, like heart and kidney, make protein extraction by freeze-thaw or mortar and pestle methods inefficient without the aid of detergent. Conversely, soft tissues, such as spleen and liver, require a milder extraction protocol for optimal results. T-PER Reagent was designed for use with all tissues, independent of mechanical extraction methods. The mild detergent enhances solubilization of proteins without compromising function, and the neutral pH and salt concentration of this reagent enable more efficient extraction from cellular compartments yielding homogeneous lysates.
T-PER Reagent was validated for protein yield in both fresh and frozen tissue. Frozen tissue and freeze-thaw extraction protocols predictably resulting in higher protein yield. Additionally, T-PER Reagent is shown to efficiently extract target proteins from various cellular compartments such as plasma membrane ( Na-K+ ATPase, pan-cadherin), cytoplasm (HSP90, MAPK), Mitochondria (COXIV), and nucleus (HDAC1) as tissue specific markers from brain (synaptophysin) and heart tissue (Troponin T Cardiac Isoform).
Related Products
M-PER™ Mammalian Protein Extraction Reagent
Features of T-PER Tissue Protein Extraction Reagent:
• Simple procedure—just homogenize tissue sample in 1:20 (w/v) of tissue to T-PER Reagent, then centrifuge to pellet cell or tissue debris
• Mild and convenient—detergent is dialyzable for quick and easy removal; can be used with additional components (e.g., protease inhibitors, salts, reducing agents, chelating agents, etc.)
• Flexible—lysate may be used for reporter assays, protein kinase assays, immunoassays and/or protein purifications
• Compatible—the lysate is compatible with standard protein assays such as Pierce Coomassie Plus (Bradford) Protein Assay
Product Details
This tissue cell lysis reagent utilizes a proprietary detergent in 25mM bicine, 150mM sodium chloride (pH 7.6) to maximize the efficiency of protein solubilization from mammalian tissue samples by homogenization. The simple composition is compatible with additives such as protease inhibitors, salts, reducing agents, chelating agents, providing versatility for many different sample types and lysis applications. Cell lysates prepared with T-PER Reagent are directly compatible with reporter assays (e.g., luciferase, beta-galactosidase, chloramphenicol acetyl transferase), protein kinase assays (e.g., PKA, PKC, tyrosine kinase), immunoassays (e.g., Western blots, ELISAs, RIAs) and/or protein purification procedures.
Homogeneous and efficient tissue protein extraction from different samples is challenging due to varying tissue composition and extraction methods. Fibrous and collagenous tissues, like heart and kidney, make protein extraction by freeze-thaw or mortar and pestle methods inefficient without the aid of detergent. Conversely, soft tissues, such as spleen and liver, require a milder extraction protocol for optimal results. T-PER Reagent was designed for use with all tissues, independent of mechanical extraction methods. The mild detergent enhances solubilization of proteins without compromising function, and the neutral pH and salt concentration of this reagent enable more efficient extraction from cellular compartments yielding homogeneous lysates.
T-PER Reagent was validated for protein yield in both fresh and frozen tissue. Frozen tissue and freeze-thaw extraction protocols predictably resulting in higher protein yield. Additionally, T-PER Reagent is shown to efficiently extract target proteins from various cellular compartments such as plasma membrane ( Na-K+ ATPase, pan-cadherin), cytoplasm (HSP90, MAPK), Mitochondria (COXIV), and nucleus (HDAC1) as tissue specific markers from brain (synaptophysin) and heart tissue (Troponin T Cardiac Isoform).
Related Products
M-PER™ Mammalian Protein Extraction Reagent
Y-PER™ Yeast Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific Y-PER Yeast Protein Extraction Reagent is the first commercially available yeast lysis reagent using a mild detergent lysis procedure to efficiently release functionally active solubilized proteins.
Features of Y-PER Yeast Protein Extraction Reagent:
• More protein—extracts more than twice as much protein as glass bead methods
• Less hassle—eliminates the difficulties associated with traditional glass bead lysis (e.g., clinging static-charged beads, protein-bead clumps and runaway beads)
• Optmized for yeast—validated for use with Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris
• Versatile—effective for many different organisms, including Bacillus subtilus and both Gram-positive and Gram-negative bacteria; suitable for use in a diverse range of situations
The detergent-based cell lysis buffer eliminates the need to use glass beads or mechanical treatments involving strong reducing agents, chemicals, pH and temperature extremes to disrupt the proteinaceous cell envelope required for protein extractions. The popular glass bead lysis protocol requires special equipment and must be performed at 4°C. The low yields of protein commonly obtained with this technique are the result of denaturation and proteins nonspecifically binding to the glass beads. However, Y-PER Reagent uses a simple room temperature protocol that can be completed in 20 minutes and requires no special equipment.
Y-PER Reagent offers an easy to use standard protocol for protein extractions involving addition of an appropriate volume of Y-PER Reagent to pelleted yeast cells, followed by incubation at room temperature for approximately 20 minutes and then spin down the debris. The resulting supernatant is a concentrated protein solution, surpassing typical yields obtained with traditional glass bead disruption.
The unicellular nature of yeast, combined with its ability to perform post-translational modifications that closely mimic processes in higher eukaryotes, have made them important research tools. The many vectors available, as well as the development of recombinant expression systems, have given yeast an irreplaceable role in the research sector.Y-PER Reagent is effective for S. cerevisiae and other popular species, making it applicable for use in fusion-tagged protein purification and reporter enzyme assays with these model organisms. The lysis reagent also can be used for genomic and plasmid DNA extraction from yeast.
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Y-PER™ Plus Dialyzable Yeast Protein Extraction Reagent
Features of Y-PER Yeast Protein Extraction Reagent:
• More protein—extracts more than twice as much protein as glass bead methods
• Less hassle—eliminates the difficulties associated with traditional glass bead lysis (e.g., clinging static-charged beads, protein-bead clumps and runaway beads)
• Optmized for yeast—validated for use with Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris
• Versatile—effective for many different organisms, including Bacillus subtilus and both Gram-positive and Gram-negative bacteria; suitable for use in a diverse range of situations
The detergent-based cell lysis buffer eliminates the need to use glass beads or mechanical treatments involving strong reducing agents, chemicals, pH and temperature extremes to disrupt the proteinaceous cell envelope required for protein extractions. The popular glass bead lysis protocol requires special equipment and must be performed at 4°C. The low yields of protein commonly obtained with this technique are the result of denaturation and proteins nonspecifically binding to the glass beads. However, Y-PER Reagent uses a simple room temperature protocol that can be completed in 20 minutes and requires no special equipment.
Y-PER Reagent offers an easy to use standard protocol for protein extractions involving addition of an appropriate volume of Y-PER Reagent to pelleted yeast cells, followed by incubation at room temperature for approximately 20 minutes and then spin down the debris. The resulting supernatant is a concentrated protein solution, surpassing typical yields obtained with traditional glass bead disruption.
The unicellular nature of yeast, combined with its ability to perform post-translational modifications that closely mimic processes in higher eukaryotes, have made them important research tools. The many vectors available, as well as the development of recombinant expression systems, have given yeast an irreplaceable role in the research sector.Y-PER Reagent is effective for S. cerevisiae and other popular species, making it applicable for use in fusion-tagged protein purification and reporter enzyme assays with these model organisms. The lysis reagent also can be used for genomic and plasmid DNA extraction from yeast.
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Y-PER™ Plus Dialyzable Yeast Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific B-PER Complete Bacterial Protein Extraction Reagent is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
Features of the B-PER Bacterial Protein Extraction Reagent:
• Detergent in Tris buffer with Lysozyme and Universal Nuclease
• Improved cell lysis and DNA digestion, thereby releasing soluble proteins and reducing viscosity to increase yields
• Recovery of large molecular weight proteins (high MW proteins)
• Cost-effective equivalent to Merck Millipore Bugbuster™ Master Mix
B-PER Complete Bacterial Protein Extraction Reagent is an all-inclusive formulation that combines the lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram-positive bacteria. Traditional Tris-buffered and phosphate-buffered formulations without enzymes are also offered. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Reagent (in Phosphate Buffer)
B-PER™ II Bacterial Protein Extraction Reagent (2X)
Features of the B-PER Bacterial Protein Extraction Reagent:
• Detergent in Tris buffer with Lysozyme and Universal Nuclease
• Improved cell lysis and DNA digestion, thereby releasing soluble proteins and reducing viscosity to increase yields
• Recovery of large molecular weight proteins (high MW proteins)
• Cost-effective equivalent to Merck Millipore Bugbuster™ Master Mix
B-PER Complete Bacterial Protein Extraction Reagent is an all-inclusive formulation that combines the lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram-positive bacteria. Traditional Tris-buffered and phosphate-buffered formulations without enzymes are also offered. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Reagent (in Phosphate Buffer)
B-PER™ II Bacterial Protein Extraction Reagent (2X)
B-PER™ Direct Bacterial Protein Extraction Kit (Thermo Scientific™)
Thermo Scientific B-PER Direct Bacterial Protein Extraction Kit is an easy-to-use cell lysis reagent is a nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.
Features of the B-PER Bacterial Protein Extraction Reagent:
• Tris buffered formulation with lysozyme and DNase enzymes
• Lysis of bacteria directly in cell culture media
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
Related Products
B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent
Features of the B-PER Bacterial Protein Extraction Reagent:
• Tris buffered formulation with lysozyme and DNase enzymes
• Lysis of bacteria directly in cell culture media
All Thermo Scientific B-PER Protein Extraction Reagents are:
• Ready to use—one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
• Fast and simple—just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
• Convenient – B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
• Excellent yields—recover recombinant proteins from bacterial lysates or purify inclusion bodies to near-homogeneous levels
• Flexible—B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
• Compatible—completely compatible with addition of protease inhibitors, and the resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in Tris or phosphate buffer at physiological pH. They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.
More Product Data
• Improved, all-in-one B-PER Reagent for bacterial protein extraction
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B-PER™ Bacterial Protein Extraction Reagent
B-PER™ Complete Bacterial Protein Extraction Reagent
I-PER™ Insect Cell Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific I-PER Insect Cell Protein Extraction Reagent enables gentle and effective extraction of soluble protein from baculovirus-infected insect cells grown in suspension or monolayer culture.
Features of I-PER Insect Cell Protein Extraction Reagent :
• Gentle extraction—optimized, mild non-ionic detergent provides maximum extraction of soluble proteins from insect cells
• Effective—provides better protein extraction than sonication
• Compatible—downstream compatibility with Western blotting, 6xHis-tagged protein purification, protein assays and ion-exchange chromatography
• Flexible—useful for protein extraction from suspended or adherent cultured insect cells
The baculovirus insect cell expression system is an efficient and popular system for the production of recombinant (eukaryotic) proteins in cell culture. Proteins expressed in baculoviral systems can be used for structural analyses, biochemical assays and a variety of other applications. I-PER Reagent maintains functionality of extracted proteins and is directly compatible with downstream applications such as protein assays, Western blotting and 6xHis-tagged protein purification.
Features of I-PER Insect Cell Protein Extraction Reagent :
• Gentle extraction—optimized, mild non-ionic detergent provides maximum extraction of soluble proteins from insect cells
• Effective—provides better protein extraction than sonication
• Compatible—downstream compatibility with Western blotting, 6xHis-tagged protein purification, protein assays and ion-exchange chromatography
• Flexible—useful for protein extraction from suspended or adherent cultured insect cells
The baculovirus insect cell expression system is an efficient and popular system for the production of recombinant (eukaryotic) proteins in cell culture. Proteins expressed in baculoviral systems can be used for structural analyses, biochemical assays and a variety of other applications. I-PER Reagent maintains functionality of extracted proteins and is directly compatible with downstream applications such as protein assays, Western blotting and 6xHis-tagged protein purification.
Y-PER™ Plus Dialyzable Yeast Protein Extraction Reagent (Thermo Scientific™)
Thermo Scientific Y-PER-Plus Dialyzable Yeast Protein Extraction Reagent effectively lyses yeast and bacterial cells to extract and solubilize proteins for analysis.
Features of Y-PER Plus Dialyzable Yeast Protein Extraction Reagent:
• Stable formulation designed for room temperature storage
• Yields protein extract with simple 20-minute hands-off incubation
• Requires no rigorous mechanical disruption or lengthy enzymatic treatments
• Tris-based buffer has a very low ionic strength and does not include sodium chloride
• Simple formulation contains no reducing agents or chelating agents
• Fully dialyzable formulation , allowing complete exchange of the extracted protein into any buffer required by downstream techniques
• Compatible with both the Thermo Scientific Pierce BCA and Coomassie Plus Protein Assays
Product Details (Recommendations for Use):
• Fresh Cells and Frozen Cells: The Y-PER-Plus Reagent is capable of extracting proteins equally well from both freshly harvested and frozen cells
• Cell Density and Strain Variation: Differences in growth rate among organisms, growth temperature and media composition affects the number of cells harvested from a given culture volume. Therefore several suggestions for the amount of Y-PER-Plus Reagent to use for a given cell pellet (wet cell paste) weight are included.
• Pichia pastoris: To achieve lysis of cells grown in rich media such as YEPD, cells must be harvested during log-phase growth. To enhance lysis of cells in stationary phase, add DTT or TCEP-HCl (to a final concentration of 0.1M and 20 to 50 mM, respectively) directly to the Y-PER-Plus Reagent before lysis to release much more of the soluble proteins. For best results use 2.5 to 5.0 mL of Y-PER-Plus Reagent per 1 gram of cell paste; scale up or down accordingly. An alternative to adding reducing agents is adding a protease inhibitor and incubating at 45°C.
• Saccharomyces cerevisiae: For best results, harvest cells in log-phase. To increase yield for cells in stationary-phase, add DTT or TCEP·HCl (to a final concentration of 0.1M and 20 to 50 mM, respectively) directly to the Y-PER-Plus Reagent before extraction. No differences in efficiency results for either rich- or synthetic-defined media. For best results use 2.5 to 5.0 mL of Y-PER Plus Reagent per 1 gram of cell paste; scale up or down accordingly.
• Schizosaccharomyces pombe: For best results, use the original Y-PER Reagent (Part No. 78990) for extracting proteins from S. pombe.
• Bacillus subtilis: The Y-PER-Plus Reagent will not extract protein from B. subtilis spores. When using a strain that is able to sporulate, harvest cells during log-phase. For strains unable to sporulate, cells may be grown to saturation before harvesting. Use 2.5 to 5.0 mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.
• Escherichia coli: Use 2.5 to 5.0 mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.
• Enzyme Activity: Because all proteins differ in structure, solubility and stability, a particular protein might not retain optimum activity in the presence of Y-PER-Plus Reagent. For example, the original Y-PER Reagent works well for assaying beta-galactosidase activity, but Y-PER-Plus Reagent slows down its kinetics. Y-PER-Plus Reagent is compatible with standard affinity-purification protocols for glutathione S-transferase (GST) and His-tagged proteins.
Related Products
Y-PER™ Yeast Protein Extraction Reagent
Features of Y-PER Plus Dialyzable Yeast Protein Extraction Reagent:
• Stable formulation designed for room temperature storage
• Yields protein extract with simple 20-minute hands-off incubation
• Requires no rigorous mechanical disruption or lengthy enzymatic treatments
• Tris-based buffer has a very low ionic strength and does not include sodium chloride
• Simple formulation contains no reducing agents or chelating agents
• Fully dialyzable formulation , allowing complete exchange of the extracted protein into any buffer required by downstream techniques
• Compatible with both the Thermo Scientific Pierce BCA and Coomassie Plus Protein Assays
Product Details (Recommendations for Use):
• Fresh Cells and Frozen Cells: The Y-PER-Plus Reagent is capable of extracting proteins equally well from both freshly harvested and frozen cells
• Cell Density and Strain Variation: Differences in growth rate among organisms, growth temperature and media composition affects the number of cells harvested from a given culture volume. Therefore several suggestions for the amount of Y-PER-Plus Reagent to use for a given cell pellet (wet cell paste) weight are included.
• Pichia pastoris: To achieve lysis of cells grown in rich media such as YEPD, cells must be harvested during log-phase growth. To enhance lysis of cells in stationary phase, add DTT or TCEP-HCl (to a final concentration of 0.1M and 20 to 50 mM, respectively) directly to the Y-PER-Plus Reagent before lysis to release much more of the soluble proteins. For best results use 2.5 to 5.0 mL of Y-PER-Plus Reagent per 1 gram of cell paste; scale up or down accordingly. An alternative to adding reducing agents is adding a protease inhibitor and incubating at 45°C.
• Saccharomyces cerevisiae: For best results, harvest cells in log-phase. To increase yield for cells in stationary-phase, add DTT or TCEP·HCl (to a final concentration of 0.1M and 20 to 50 mM, respectively) directly to the Y-PER-Plus Reagent before extraction. No differences in efficiency results for either rich- or synthetic-defined media. For best results use 2.5 to 5.0 mL of Y-PER Plus Reagent per 1 gram of cell paste; scale up or down accordingly.
• Schizosaccharomyces pombe: For best results, use the original Y-PER Reagent (Part No. 78990) for extracting proteins from S. pombe.
• Bacillus subtilis: The Y-PER-Plus Reagent will not extract protein from B. subtilis spores. When using a strain that is able to sporulate, harvest cells during log-phase. For strains unable to sporulate, cells may be grown to saturation before harvesting. Use 2.5 to 5.0 mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.
• Escherichia coli: Use 2.5 to 5.0 mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.
• Enzyme Activity: Because all proteins differ in structure, solubility and stability, a particular protein might not retain optimum activity in the presence of Y-PER-Plus Reagent. For example, the original Y-PER Reagent works well for assaying beta-galactosidase activity, but Y-PER-Plus Reagent slows down its kinetics. Y-PER-Plus Reagent is compatible with standard affinity-purification protocols for glutathione S-transferase (GST) and His-tagged proteins.
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Y-PER™ Yeast Protein Extraction Reagent
Total Exosome RNA & Protein Isolation Kit (Invitrogen™)
The Total Exosome RNA and Protein Isolation Kit enables isolation of total RNA and proteins from exosome samples extracted from biological fluids (e.g., by using Total Exosome Isolation (from cell culture media) or Total Exosome Isolation (from serum)).
• Extracts highly pure total RNA (including small RNA fraction) from exosomes
• Recovers protein and RNA from the same sample
• Compatible with all protocols for exosome isolation
The Total Exosome RNA and Protein Isolation Kit is designed for the isolation of both RNA and protein from pre-isolated or enriched exosomes (not supplied). It is suitable for studies of RNA expression (specifically miRNA), processing, or function. The kit allows recovery of both RNA and proteins from the same purified exosome preparation. One portion of the sample is subjected to organic extraction, followed by immobilization of RNA on glass-fiber filters to purify total RNA or enrich for small RNA species. The other portion is then used directly for common protein applications such as Western blotting.
High yields of ultra-pure RNA can be prepared in about 30–60 minutes. The RNA can then be used in a variety of downstream applications, including qRT-PCR, high-throughput sequencing (e.g., using the Ion Torent™ PGM™, Ion Torrent™ Proton™, or SOLiD® sequencing instruments), microarray analysis, solution hybridization assays, and blot hybridization.
For Research Use Only. Not for human or animal therapeutic or diagnostic use.
• Extracts highly pure total RNA (including small RNA fraction) from exosomes
• Recovers protein and RNA from the same sample
• Compatible with all protocols for exosome isolation
The Total Exosome RNA and Protein Isolation Kit is designed for the isolation of both RNA and protein from pre-isolated or enriched exosomes (not supplied). It is suitable for studies of RNA expression (specifically miRNA), processing, or function. The kit allows recovery of both RNA and proteins from the same purified exosome preparation. One portion of the sample is subjected to organic extraction, followed by immobilization of RNA on glass-fiber filters to purify total RNA or enrich for small RNA species. The other portion is then used directly for common protein applications such as Western blotting.
High yields of ultra-pure RNA can be prepared in about 30–60 minutes. The RNA can then be used in a variety of downstream applications, including qRT-PCR, high-throughput sequencing (e.g., using the Ion Torent™ PGM™, Ion Torrent™ Proton™, or SOLiD® sequencing instruments), microarray analysis, solution hybridization assays, and blot hybridization.
For Research Use Only. Not for human or animal therapeutic or diagnostic use.
Pierce™ Cell Surface Protein Isolation Kit (Thermo Scientific™)
The Thermo Scientific Cell Surface Protein Isolation Kit is a complete set of reagents for selective biotinylation and subsequent purification of mammalian cell surface proteins to the exclusion of intracellular proteins.
Features of the Cell Surface Protein Isolation Kit:
• Isolates cell surface proteins—reduces complexity of total cellular protein
• Efficiently recovers labeled proteins—cleavable biotin allows for nearly 100% recovery of isolated cell surface proteins
• Convenience—all reagents are provided in one kit, along with complete instructions for labeling, cell lysis and purification of cell surface membrane proteins
• Western blotting applications—proteins recovered in SDS-PAGE buffer are loaded directly onto polyacrylamide gels
• Robust system—protocol designed for diverse cell lines, including NIH 3T3, HeLa, C6 and A431
The kit efficiently labels proteins that have accessible lysine residues and sufficient extracellular exposure. The isolation procedure uses a cell-impermeable, cleavable biotinylation reagent (Sulfo-NHS-SS-Biotin) to label exposed primary amines of proteins on the surface of intact adherent or suspension cells. Treated cells are then harvested, lysed and the labeled surface proteins are affinity-purified using Thermo Scientific NeutrAvidin Agarose Resin. The isolated cell surface proteins contain a small, nonreactive tag of the originally labeled primary amines but are no longer biotinylated (biotin remains bound to the resin). The kit contains sufficient reagents for eight experiments, each involving four T75 flasks of confluent cells.
The Cell Surface Protein Isolation Kit has been successfully used to label and purify cell surface proteins. In one experiment, HeLa cells were treated with or without Sulfo-NHS-SS-Biotin and the cells were processed according to the kit procedure. Fractions collected, including elution, flow-through and post-elution gel, were analyzed by Western blot. Four cell surface proteins were identified in the elution fraction of cells treated with the biotin label. These proteins included epidermal growth factor receptor (EGFR), insulin-like growth factor-1 receptor beta (IGF-1β), integrin 1 and integrin 5. In marked contrast, none of these proteins were identified in the elution fraction of unlabeled cells; the unlabeled proteins were exclusively found in the flow-through. Some cell surface proteins were also identified in the flow-through fractions from labeled cells. These proteins were likely not labeled because of steric hindrance. A negligible amount of captured protein was present in the post-elution gel fractions, indicating nearly 100% recovery of captured proteins. A trace amount of heat shock protein 90 (Hsp90) was detected in the elution fraction of all samples tested, demonstrating the specificity of the label for cell surface versus intracellular proteins.
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Subcellular Protein Fractionation Kit for Cultured Cells
Subcellular Protein Fractionation Kit for Tissues
Features of the Cell Surface Protein Isolation Kit:
• Isolates cell surface proteins—reduces complexity of total cellular protein
• Efficiently recovers labeled proteins—cleavable biotin allows for nearly 100% recovery of isolated cell surface proteins
• Convenience—all reagents are provided in one kit, along with complete instructions for labeling, cell lysis and purification of cell surface membrane proteins
• Western blotting applications—proteins recovered in SDS-PAGE buffer are loaded directly onto polyacrylamide gels
• Robust system—protocol designed for diverse cell lines, including NIH 3T3, HeLa, C6 and A431
The kit efficiently labels proteins that have accessible lysine residues and sufficient extracellular exposure. The isolation procedure uses a cell-impermeable, cleavable biotinylation reagent (Sulfo-NHS-SS-Biotin) to label exposed primary amines of proteins on the surface of intact adherent or suspension cells. Treated cells are then harvested, lysed and the labeled surface proteins are affinity-purified using Thermo Scientific NeutrAvidin Agarose Resin. The isolated cell surface proteins contain a small, nonreactive tag of the originally labeled primary amines but are no longer biotinylated (biotin remains bound to the resin). The kit contains sufficient reagents for eight experiments, each involving four T75 flasks of confluent cells.
The Cell Surface Protein Isolation Kit has been successfully used to label and purify cell surface proteins. In one experiment, HeLa cells were treated with or without Sulfo-NHS-SS-Biotin and the cells were processed according to the kit procedure. Fractions collected, including elution, flow-through and post-elution gel, were analyzed by Western blot. Four cell surface proteins were identified in the elution fraction of cells treated with the biotin label. These proteins included epidermal growth factor receptor (EGFR), insulin-like growth factor-1 receptor beta (IGF-1β), integrin 1 and integrin 5. In marked contrast, none of these proteins were identified in the elution fraction of unlabeled cells; the unlabeled proteins were exclusively found in the flow-through. Some cell surface proteins were also identified in the flow-through fractions from labeled cells. These proteins were likely not labeled because of steric hindrance. A negligible amount of captured protein was present in the post-elution gel fractions, indicating nearly 100% recovery of captured proteins. A trace amount of heat shock protein 90 (Hsp90) was detected in the elution fraction of all samples tested, demonstrating the specificity of the label for cell surface versus intracellular proteins.
Related Products
Subcellular Protein Fractionation Kit for Cultured Cells
Subcellular Protein Fractionation Kit for Tissues
