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Genomics and Proteomics


Column Extenders for Pierce™ Centrifuge Columns Thermo Scientific™

Thermo Scientific Pierce Centrifuge Column Extenders are made of polypropylene, and are designed for use in gravity-flow procedures. They fit 2 mL, 5 mL and 10 mL Pierce Centrifuge Columns.

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Pierce™ Centrifuge Columns, 0.8 mL
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Pierce™ Centrifuge Columns, 5 mL
Pierce™ Centrifuge Columns, 10 mL

Pierce™ Microcentrifuge Tubes, 1.5 mL Thermo Scientific™

Thermo Scientific Pierce 1.5 mL Capacity Microcentrifuge Tubes are convenient tools for manipulating small volumes of affinity supports for protein purification.

Features of 1.5 mL Capacity Microcentrifuge Tubes:

• 1.5 mL capacity
• Polypropylene formulation
• Attached snap caps

Simply add the affinity resin and sample to one of the microcentrifuge tubes, then use a microcentrifuge to efficiently wash away contaminants and elute your purified sample without losing any resin in the process. Microcentrifuge tubes allow you to affinity purify more protein in less time.

Applications:
• Affinity purification or affinity chromatography
• Immunodepletion
• Immunoprecipitation (IP)
• Co-immunoprecipitation (co-IP)

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Pierce™ Microcentrifuge Tubes, 2.0 mL

Pierce™ Microcentrifuge Tubes, 2.0 mL Thermo Scientific™

Thermo Scientific Pierce 2 mL Capacity Microcentrifuge Tubes are convenient tools for manipulating small volumes of affinity supports for protein purification.

Features of 2 mL Capacity Microcentrifuge Tubes:

• 1.5 mL capacity
• Polypropylene formulation
• Attached snap caps

Simply add the affinity resin and sample to one of the microcentrifuge tubes, then use a microcentrifuge to efficiently wash away contaminants and elute your purified sample without losing any resin in the process. Microcentrifuge tubes allow you to affinity purify more protein in less time.

Applications:
• Affinity purification or affinity chromatography
• Immunodepletion
• Immunoprecipitation (IP)
• Co-immunoprecipitation (co-IP)

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Pierce™ Microcentrifuge Tubes, 1.5 mL

Pierce™ NHS-Activated Agarose Slurry Thermo Scientific™

Thermo Scientific Pierce NHS-Activated Agarose Slurry is a high-quality, amine-reactive, beaded-agarose resin for rapid and stable immobilization of proteins, peptides and other ligands via primary amines.

Features of NHS-Activated Agarose:

Easy to use—immobilize in a simple one-step reaction with minimal hands-on time
Rapid and efficient—greater than 85% coupling for most proteins within 30 minutes
Innovative format—the dry agarose concentrates the sample, making it ideal for immobilizing dilute proteins
Safe—No hazardous chemicals needed (e.g., sodium cyanoborohydride, cyanogen bromide)
Versatile—affinity resin is adaptable to column and batch affinity chromatography techniques and FPLC applications
Compatible—use with any primary amine-containing compound
Reusable—the leak-resistant chemistry means you can reuse the affinity resin
High binding capacity—coupling capacity of >30 mg/mL

NHS-activated agarose is crosslinked, 6% beaded agarose resin that contains N-hydroxysuccinimide (NHS) functional groups. The activated resin reacts with primary amines to form stable amide linkages that covalently immobilize antibodies or other proteins for use in affinity purification procedures. Pierce NHS-Activated Agarose has a coupling capacity greater than 30 mg/mL for the slurry.

Applications:
• Rapid Immobilization of goat anti-mouse and anti-rabbit antibodies in order to purify IgG produced in animals or hybridomas.
• Bulk immobilization of Protein A for purification of monoclonal antibodies
• Immobilization of ligands for purification of recombinant proteins

Pierce NHS-Activated Agarose resin uses reliable NHS-ester Chemistry and does not require hazardous chemicals for immobilization. Other amine-reactive supports, such as periodate-oxidized resins, use toxic sodium cyanoborohydride to stabilize the reaction linkage to primary amines and take 4 to 6 hours to complete. Traditional methods such as cyanogen bromide-activated supports also couple amines; however, this chemistry results in nonspecific binding and constant slow leakage of the coupled ligand. Reactions with Pierce NHS-Activated Agarose are complete in less than one hour and yield much more stable linkages.

The NHS-Activated Agarose coupling reaction is performed in an amine-free buffer at pH 7-9. Protein coupling efficiency is typically greater than 80%, regardless of the ligand's molecular weight or pI. Once a ligand is immobilized, the prepared resin can be used for multiple affinity purification procedures. The crosslinked beaded agarose has fast linear flow potential, making it useful for gravity-flow and low- to medium-pressure applications.

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Pierce™ NHS-Activated Agarose, Dry
Pierce™ NHS-Activated Agarose Spin Columns, 2 mL
Pierce™ NHS-Activated Agarose Spin Columns, 0.2 mL

Pierce™ Spin Columns - Snap Cap Thermo Scientific™

Thermo Scientific Pierce Snap Cap Spin Columns are convenient tools for manipulating small volumes of affinity supports (20 to 500 µL) for protein purification.

Features of Snap Cap Spin Columns:

Formulation: polypropylene
Column volume: 1 mL
Resin volume: 20 to 500µL
Filter type: Polyethylene filter, ~30 µm pore size
Cap: Attached snap-cap (no cap on collection tube); press-on bottom caps

Simply add the affinity resin and sample to one of the spin columns, then use a microcentrifuge to efficiently wash away contaminants and elute your purified sample without losing any resin in the process. Spin columns allow you to affinity purify more protein in less time.

Applications:
• Affinity purification or affinity chromatography
• Immunodepletion
• Immunoprecipitation (IP)
• Co-immunoprecipitation (co-IP)

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Pierce™ Monomeric Avidin Agarose Thermo Scientific™

Thermo Scientific Pierce Monomeric Avidin Agarose is ideal for purifying biotinylated proteins, peptides and other molecules.

Features of Monomeric Avidin Agarose:

Non-denaturing—purifies biotinylated products using mild elution conditions (2 mM free biotin)
Reusable—Monomeric Avidin Agarose can be regenerated and reused at least 10 times, with only a marginal loss in biotin binding capacity (approximately 2.5% decrease per regeneration)
Specific—retains biotin-binding specificity and low nonspecific binding of native avidin
Flexible—bind samples in a variety of physiological buffers and elute either with 0.1M glycine or by competition with 2 mM biotin
Good binding capacity—greater than 1.2 mg biotinylated BSA/mL resin

The Immobilized Monomeric Avidin protein binds biotin with high specificity and moderate affinity, allowing non-biotin molecules to be washed away and then the bound biotin-labeled molecules to be competitively eluted using 2 mM biotin in phosphate buffered saline (PBS). This technique provides the gentlest elution conditions for biotinylated protein purification and avoids the contamination and other problems associated with traditional avidin and streptavidin methods. The monomeric avidin column can then be regenerated by using 0.1M glycine to strip the column of residual bound biotin without losing the ability to bind another biotinylated sample.

With typical avidin or streptavidin, the biotin-binding affinity (Kd = 10-15M) is so great that purification with these traditional media require denaturing conditions for elution, such as 8M Guanidine·HCl at pH 1.5 or boiling in reducing SDS-PAGE sample loading buffer. In addition to their adverse effects on the biotinylated protein of interest, such harsh elution conditions typically cause extraneous proteins (i.e., the avidin or streptavidin subunits) to strip from the resin and co-elute with the desired purification product. By contrast, when avidin is coupled to a resin (e.g., crosslinked beaded agarose or UltraLink Resin) as the subunit monomer, its specificity for biotin is retained, but its biotin-binding affinity decreases to a level (Kd = 10-8M) that is conducive to subsequent elution.

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Pierce™ Monomeric Avidin Agarose Kit, 2 mL
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POROS™ MC 20 µm Column, 2.1 x 30 mm, 0.1 mL Thermo Scientific™

Applied Biosystems™ POROS™ Prepacked Immobilized Metal Affinity Column. POROS™ MC is based on a Imido-diacetate functional group designed for the purifiction of proteins that form co-ordination complexes via metal sites and specific amino acids, such as histidine tagged fusion proteins.
20 micron particle size is used for high resolution and small scale preparative to semi-preparative separation of biomolecules.

POROS™ High Performance Chromatography™ Media and Pre-Packed Columns

A high performance chromatography resins for analytical to process scale separations.• Higher productivity: High throughput and high dynamic capacity associated with High Performance Chromatography™
• Chemical stability: Allows aggressive cleaning and sanitization
• Enhanced biomolecule access: Provided via large pores, ranging between 500-10000 Å
• Polystyrenedivinylbenzene particles: Yield a robust, easily packable matrix
• Develop better separations methods in a shorter time frame: The speed of High Performance Chromatography technology reduces weeks of experimentation to only a few hours of work, so you have plenty of time to explore all the variables of your separation

Reduce time-consuming sample prep

You can replace many sample preparation steps with high speed High Performance Chromatography technology. For instance, dialysis of large volumes of material can be replaced by high flow rate processing of the dilute sample. Elution in a small volume of new buffer accomplishes both buffer exchange and concentration.

Create novel assays for more efficient analysis

High Performance Chromatography technology is not limited to standard modes of separation. Both enzymes and affinity ligands can be immobilized on POROS media. By combining rapid on-column protein digestions with rapid on-column immunoassays and chromatographic separations, you can create entirely new assays with unlimited potential.

High capacity, high resolution, high speed
In contrast to conventional chromatography media, POROS™ High Performance Chromatography™ resins particles, are engineered to have two discreet classes of pores. Large "throughpores" allow convection flow to occur through the particles themselves, quickly carrying sample molecules to short "diffusive" pores inside. By reducing the distance over which diffusion needs to occur, the time required for sample molecules to interact with interior binding sites is also reduced. Diffusion is no longer limiting and flow rates can be dramatically increased - without compromising resolution or capacity. Separations can be achieved at 1,000 to 5,000 cm⁄hr compared to 50 to 360 cm⁄hr for conventional media.

Pierce™ Anti-c-Myc Agarose Thermo Scientific™

Thermo Scientific Pierce Anti-c-Myc Agarose is a high-affinity immunoprecipitation resin ideal for purification of recombinant c-Myc-tagged proteins expressed in human in vitro expression systems and bacterial and mammalian cell lysates.

Features of Anti-c-Myc Agarose:

Specific—highly specific anti-c-Myc monoclonal antibody (clone 9E10) enables high yield and high purity for immunoprecipitation
Scalable—available in 2 and 10 mL resin package sizes to allow for larger scale purifications or immunoprecipitations
Versatile—can be used in spin or gravity columns as well as in FPLC cartridges
Convenient—reagents to elute and detect c-Myc tagged fusion proteins are available separately

The anti-c-Myc antibody used to manufacture Pierce Anti-c-Myc Agarose is a highly specific mouse IgG1 monoclonal antibody (clone 9E10) that recognizes the c-Myc epitope tag (EQKLISEEDL) derived from the human c-myc oncogene (p62 c-myc). The support is Superflow 6 resin, a highly crosslinked 6% agarose resin. This anti-c-Myc affinity resin can be packed into gravity purification columns, spin purification columns or cartridges for FPLC instruments to purify c-Myc-fusion proteins expressed in bacterial or mammalian cells.

Applications:
• Purification of c-Myc fusion proteins expressed in the Pierce Human In Vitro Translation kits
• Large scale purification of recombinant c-Myc tagged proteins
• High-throughput enrichment of fusion proteins and interacting partners
• IP and co-IP experiments (as with the Pierce c-Myc-Tag IP/co-IP Kit)

Pierce Anti-c-Myc Agarose consists of a highly specific monoclonal anti-c-Myc antibody that is covalently immobilized on a crosslinked 6% beaded agarose support. The product is supplied as a 25% slurry. Upon incubation with a sample, c-Myc tagged fusion protein is captured on the agarose beads. After simple washing steps, the tagged protein is easily eluted from the resin using 0.1M glycine (pH 2.0-2.8), 3M NaSCN or 50 mM NaOH depending on the downstream application of the purified protein. For elution of highly functional proteins, Pierce c-Myc-peptide can also be used to elute the c-Myc tagged protein. Anti-c-Myc antibody may be used to detect the presence of the tagged protein by Western blot.

In experiments with a c-Myc-tagged fusion protein (26 to 29kDa), the resin provided a binding capacity of 102 to 144nmol protein per mL of settled resin. The elution capacity was at least 18 to 19nmol per mL of settled resin using 0.1M glycine, pH 2.8. Binding and elution capacity will vary depending on the c-Myc-fusion protein and the method of elution.

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CaptureSelect™ C-tagXL Affinity Matrix Thermo Scientific™

CaptureSelect C-tagXL Affinity Matrix combines a unique selectivity for a small 4-amino acid peptide tag (E-P-E-A, glutamic acid-proline-glutamic acid-alanine) with the benefits of a robust and high quality affinity matrix. It purifies C-terminal tagged proteins with high affinity and selectivity, even in the presence of urea and guanidine HCl, from complex mixtures like cell culture harvests and periplasmatic fractions in a one-step process. Mild elution conditions at neutral pH can be applied using magnesium chloride or propylene glycol, which ensures high activity recoveries of pH-sensitive target proteins. C-tagXL Affinity Matrix recognizes the E-P-E-A tag sequence when fused either directly to the C-terminus of a protein or through a linker between the C-terminus and the E-P-E-A tag.

Features of C-tagXL Affinity Matrix include:
• Mild elution, making it suitable for pH-sensitive proteins
• Binding of the tagged proteins under denaturing conditions (like dissolved inclusion bodies)
• Excellent scalability
• Non-animal-derived

CaptureSelect C-tagXL Affinity Matrix purifies C-terminal tagged E-P-E-A proteins directly from complex source materials in a single step with high purity and yield.

Free of animal components
Our CaptureSelect products are affinity ligands created by a proprietary technology based on camelid-derived single-domain antibody fragments. The ligand is a 13-kDa fragment comprising the three complementarity-determining regions (CDRs) that form the antigen-binding domain, efficiently produced by the yeast Saccharomyces cerevisiae in a production process free of animal components.

Main characteristics:

Matrix: agarose-based, epoxide-activated
Average particle size: 65 ± 10 µm
Ligand: CaptureSelect C-tagXL affinity ligand
Ligand coupling method: epoxide coupling of the ligand
Binding capacity: 400 nmol/mL resin depending on flow rate, column height, and contact time
Elution conditions: Acidic: 20 mM citric acid or acetic acid, pH 3–4, or 0.1 M glycine pH 3.0; Neutral: 20 mM Tris, 2.0 M MgCl2 pH 7; 20 mM Tris, 1.0 M NaCl, 50% (v/v) propylene glycol; 20 mM Tris 2 mM S-E-P-E-A.
Flow characteristics: 150–300 cm/h
Formulation buffer: 20%(v/v) ethanol

n-Dodecyl-beta-Maltoside Detergent Thermo Scientific™

Thermo Scientific n-Dodecyl-β-D-Maltoside is especially useful for solubilizing membrane proteins to preserve their activity.

The water soluble nonionic detergent is most often used for the isolation of hydrophobic membrane proteins. Multiple studies have shown that n-Dodecyl-β-D-maltoside is a gentle detergent that is often able to preserve protein activity better than many commonly used detergents, including NP-40, CHAPS and Octyl-β-glucoside.

Features of n-Dodecyl-β-D-Maltoside:

• Lipid-like nonionic detergent
• Especially useful for isolating and stabilizing hydrophobic membrane proteins
• Preserves activity of membrane protein better than most of the detergents
• High-purity compound with low UV absorptivity

Like most detergents, n-Dodecyl-β-D-maltoside has dual hydrophobic/hydrophilic properties that facilitate lipid displacement and provide a lipid-like environment for membrane proteins. Studies suggest that the ability of these surfactants to preserve membrane protein structure stems in part from the reduced disruption of lipid:protein interactions where some of the natural lipid associations are maintained. While no detergent or set of conditions is optimal for all studies on membrane proteins, n-Dodecyl-β-D-maltoside has unique characteristics that have made it especially helpful in certain protein methods.

Properties of n-Dodecyl-β-D-Maltoside:
• Chemical Name: n-Dodecyl-beta-D-maltoside
• Molecular Weight: 510.6g
• Detergent Class: Nonionic
• Aggregation Number: 98 (average), 70 to 140 range
• Micelle Molecular Weight: 50,000g
• Critical Micelle Concentration (CMC): 0.17 mM (0.009%, w/v) in water ; 0.12 mM (0.006%, w/v) in 0.2M NaCl
• Cloud Point: Unknown
• Dialyzable: No

Specifications for n-Dodecyl-beta-maltoside (Part No. 89902, 89903):
• Formula: C24H46O11
• Molecular Weight: 510.6g
• Purity (by HPLC): ≥99%
• Absorbance (1% Detergent Solution): 340nm <0.02; 280nm <0.04; 260nm <0.06; 225nm <0.1
• pH (1% Solution): 5 to 8
• Solubility (in water at 0 to 5°C): ≥20%
• Conductivity (10% Solution) : <40mS

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CaptureSelect™ Alpha-1 Antitrypsin Affinity Matrix Thermo Scientific™

This is one of a group of CaptureSelect™ protein purification affinity resins developed for the simple, single-step purification of a variety human plasma and serum proteins. This unique selection of affinity purification resins can purify proteins or other non-antibody biomolecules from either recombinant or native sources with a high degree of affinity and specificity, and are immobilized on a high capacity agarose support.

POROS™ Oligo (dT)25 Affinity Resin Thermo Scientific™

POROS Oligo (dT)25 Affinity Resin is designed for the purification and isolation of mRNA from in vitro transcription (IVT) manufacturing processes. The resin effectively separates mRNA from components of the transcription reaction process, such as enzymes and plasmid DNA.

POROS Oligo (dT)25 Affinity Resin was developed to address the selectivity and capacity requirements for large-scale downstream purification of a range of mRNA constructs used in vaccine and gene therapy applications. Unlike alternate approaches, the resin selectively captures the mRNA via the polyadenylated (polyA) tail using simple salt and water purification steps.

Features of POROS Oligo (dT)25 Affinity Resin include:
• Easy mRNA purification from crude transcription mix to effectively remove plasmid DNA and other IVT components
• Simplified workflow helps to maximize efficiency, thereby reducing complexity of subsequent polish steps
• Excellent scalability
• Non-animal derived

POROS Oligo (dT)25 Affinity Resin allows for a robust platform and efficient purification process for a range of mRNA molecules and applications.

Pack sizes of 1 L and above come with a regulatory support package. Worldwide technical support is also available.

Main characteristics

Matrix: cross-linked poly(styrene-divinylbenzene) POROS beads
Average particle size: 50 µm
Ligand: poly(dT) 25mer with proprietary linker
Binding capacity: >2 mg mRNA per mL of resin
Binding conditions: 0.5–1 M salt, common buffers, neutral pH
Elution conditions: Low salt, common buffers, neutral pH
Mechanical resistance: 100 bar (1,450 psi; 10 MPa)
Thermal stability: allows sample denaturing at 65°C if needed
Formulation buffer: 18% (v/v) ethanol

Oligo (dT) ligands are manufactured using a synthetic manufacturing process that are free of animal components.

Mouse (CD-1) Cytosol Gibco™

Liver cytosolic fractions prepared from a pool of male CD-1 mice.

Total protein concentration: 20 mg/ml

We provide liver subcellular fractions from a variety of tox species, including human, nonhuman primates (Cynomolgus Monkey and Rhesus Monkey), dog (Beagle), rat (Sprague-Dawley), mouse (CD-1 and Balb-c), and trout (Oncorhynchus mykiss). Custom species are available upon request.

For research use only. Not intended for human or animal therapeutic or diagnostic use.

Related Link

Learn more about our microsomes and subcellular fractions

Rat (Sprague Dawley) Microsomes Gibco™

Liver microsomes prepared from a pool of male Sprague Dawley rats.

Total protein concentration: 20 mg/ml

We provide liver subcellular fractions from a variety of tox species, including human, nonhuman primates (Cynomolgus Monkey and Rhesus Monkey), dog (Beagle), rat (Sprague-Dawley), mouse (CD-1 and Balb-c), and trout (Oncorhynchus mykiss). Custom species are available upon request.

For research use only. Not intended for human or animal therapeutic or diagnostic use.

Related Link

Learn more about our microsomes and subcellular fractions

Pierce™ High Capacity Streptavidin Chromatography Cartridges, 1 mL Thermo Scientific™

The Thermo ScientificTM PierceTM High Capacity Streptavidin Chromatography Cartridges are convenient, ready-to-use devices for separating biotinylated molecules from non-biotinylated molecules and for purifying antigens using biotinylated antibodies. The chromatography cartridges contain a beaded resin of immobilized recombinant streptavidin protein that binds biotinylated proteins at greater than 10 mg/mL of beads. This corresponds to 2 to 3 times higher biotin-binding capacity than products from other suppliers and is ideal when a high amount of recovery is required.

These cartridges enable fast, easy, and reproducible chromatographic separations and are compatible with the major automated liquid-chromatography systems or manual syringe processing (see Table 1 for cartridge general properties). The cartridges attach directly to ÄKTATM or FPLC systems without additional connectors. The included accessory pack readily adapts cartridges for use with Luer-LokTM syringe fittings or 1/16' tubing. These cartridges can be regenerated multiple times for affinity purification.

Features of High Capacity Streptavidin Agarose:
Streptavidin—purified recombinant streptavidin protein (53 kDa, near-neutral pI); tetrameric with four biotin-binding site per molecule
Agarose resin—support is cross-linked 6% beaded agarose (CL-6B), the most popular resin for protein affinity purification methods
Inert and stable—streptavidin is immobilized by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
High capacity—this variety of beads has a dense load of immobilized streptavidin, providing a binding capacity greater than 10 mg of biotinylated BSA per mL of resin
Two formats—available in several bottle sizes of resin slurry and as packed 1 mL and 5 mL chromatography cartridges

Properties of cross-linked 6% beaded agarose (CL-6B):
• Support pH stability: 2 to 14 (short term); 3 to 13 (long term)
• Average particle size: 45 to 165 microns
• Exclusion limit: 10,000 to 4,000,000 daltons
• Maximum recommended flow rate: 4 mL/minute (1 mL column), 5 mL/min (5 mL column)
• Maximum linear velocity: 30 cm per hour
• Maximum pressure: 0.3 MPa, 43.5 psi or 3 bar

Pierce High Capacity Streptavidin Chromatography Cartridges consist of purified recombinant streptavidin covalently immobilized at a high density onto high-quality cross-linked 6% beaded agarose. Biotinylated antibodies, proteins, peptides, nucleic acids, and other molecules or interaction complexes can be captured, immunoprecipitated, removed, or purified from samples using this streptavidin resin.

Streptavidin is a tetrameric protein containing four biotin-binding sites similar to avidin. The native protein from Streptomyces avidinii is carbohydrate-free, has an acidic pI of 5.5 and a mass of 75 kDa. The recombinant protein used to manufacture Pierce streptavidin resins has a near-neutral isoelectric point and a mass of 53 kDa. Streptavidin generally has less nonspecific binding than avidin (from chicken egg white) because of the absence of carbohydrates.

For more information see: Chromatography Cartridges for Desalting and Affinity Purification

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