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Pierce™ Protein A/G Magnetic Beads Thermo Scientific™

Thermo Scientific Pierce Protein A/G Magnetic Beads are high-performance affinity particles for antibody purification and immunoprecipitation methods using manual or robotic magnetic separators.

Features of Protein A/G Magnetic Beads:

High capacity—nearly four times higher binding capacity than typical magnetic beads from other suppliers, allowing the use of smaller amounts per experiment
Low non-specific binding—stable, pre-blocked beads provide clean purification products (e.g., antigen eluted in IP with antibody is devoid of contaminating proteins from complex IP matrix)
Flexibility—convenience of IgG binding domains of both Protein A and Protein G on one bead
Compatibility—beads are compatible with manual and automated applications (e.g., Thermo Scientific KingFisher Instruments)
Assay consistency—magnetic beads eliminate resin loss and provide for more efficient separation of solutions than traditional IP methods that use only microcentrifuge tubes

These magnetic beads are coated with genetically engineered Pierce Protein A/G, a recombinant fusion protein which combines the IgG binding domains of both Protein A and Protein G. This enables capture of antibodies from a wider range of species and isotypes than either protein alone. Using our crosslinker chemistry, you can immobilize an antibody onto the magnetic particle and prevent IgG contamination in your immunoprecipitated sample. These beads can be used both manually with a magnetic stand as well as with automated platforms such as the Thermo Scientific KingFisher Instruments.

Applications:
• IP and Co-IP experiments (see complete kit)
• Immunoprecipitation for analysis in non-reducing conditions
• Antibody purification

The recombinant Protein A/G that is immobilized onto the Pierce Magnetic Beads is a fusion of the IgG binding domains of both Protein A and Protein G. Protein A/G contains four Fc-binding domains from Protein A and two from Protein G, making it a convenient tool for investigating and purifying immunoglobulins. Thus, Pierce Magnetic Particles are not simply a mixed immobilization of separate Protein A and Protein G polypeptides, nor are they a mixture of Protein A magnetic beads and Protein G magnetic beads.

Pierce™ Classic Magnetic IP/Co-IP Kit Thermo Scientific™

The Thermo Scientific Pierce Classic Magnetic IP/Co-IP Kit enables highly effective and efficient immunoprecipitation (IP) and co-immunoprecipitation (co-IP) of antigens and protein-complexes using less than 10 µg of antibody and a magnetic separator.

Features of the Classic Magnetic IP/Co-IP Kit:

Compatible—magnetic beads based on Protein A/G recombinant protein ensures compatibility with most primary antibodies, whether from mouse or rabbit
Fast—immunoprecipitate in as few as 30 minutes to reduce background and improve the capture of transient protein complexes
Clean—immobilize your antibody to prevent contamination in your eluate
Resistant—no leaching of Protein A/G in the presence of detergents, low pH buffers or common mass spectrometry solvents
Efficient—immunoprecipitate with half the recommended volume of magnetic particles compared to other magnetic beads
Convenient—the comprehensive Co-IP kit contains the magnetic beads and all essential buffers to complete immunoprecipitation experiments

This immunoprecipitation kit uses high-quality Thermo Scientific Pierce Protein A/G Magnetic Beads and optimized buffers and flexible protocol to accomplish high-yield IP or co-IP with manual or automated magnetic-separation tools. The optimized Lysis/Wash Buffer optimizes yield and efficient binding of antibody-antigen and co-IP interactions. The relatively gentle, low-pH Elution Buffer dissociates the bound immune complex from the Protein A/G. Alternatively, the included Lane Marker Sample Buffer provides rapid, denaturing elution for direct SDS-PAGE analysis of IP products.

Includes:
Pierce Protein A/G Magnetic Beads, lysis/wash buffer, elution buffer, neutralization buffer and sample loading buffer

Immunoprecipitation with magnetic particles is performed much like IP with beaded agarose, except that separations are performed using a magnet rather than by centrifugation. The specific antibody is first added to the sample to form an immune complex that is then bound to the magnetic beads. The complex is washed to remove non-bound material and a low-pH elution buffer dissociates the bound immune complex from the Protein A/G. Alternatively, the Lane Marker Sample Buffer is included for dissociation using denaturing conditions or for downstream sample prep for SDS-PAGE analysis. The kit includes Thermo Scientific Pierce Protein A/G Magnetic Beads for fast and convenient magnetic isolation of antigens and optimized buffers for high antigen yield. The beads are removed from the solution manually using a magnetic stand or by automation with an instrument such as the Thermo Scientific KingFisher Flex Instrument.

It is generally known that longer antibody-sample incubations times (2 hours to overnight) usually provide greater yields in IP assays. It is often assumed that this greater yield comes with increased background. Our researchers have found that overnight antibody-sample binding followed by immunoprecipitation with the Pierce Classic Magnetic IP/Co-IP Kit does tend to increase yield but does not increase background. Thus, we recommend that researchers perform this binding step for at least 1 hour whenever possible.

Protocol Summary:
• Incubate cell lysate with IP antibody for 1 to 2 hours at room temperature or overnight at 4ºC.
• Bind antigen-antibody complex to Protein A/G magnetic beads for 1 hour at room temperature.
• Wash beads twice with IP Lysis/Wash Buffer and once with purified water.
• Elute the antigen/antibody complex.

Related Products
Pierce™ Classic IP Kit

Alexa Fluor™ Plus 405 Phalloidin Invitrogen™

Alexa Fluor Plus 405 Phalloidin is a high-affinity filamentous actin (F-actin) probe (phalloidin) conjugated to our bright, photostable, violet-fluorescent Alexa Fluor 405 dye.

• Selectively stains F-actin
• Outstanding fluorescence performance
• Excitation/emission: 405/450 nm. Compatible with standard DAPI filter set or 405/violet excitation laser.
• Superior to antibody staining
• Optimal for fixed and permeabilized samples

Get superior results in your actin staining studies
Phalloidin is a bi-cyclic peptide belonging to a family of toxins isolated from the deadly Amanita phalloides ''death cap'' mushroom and commonly used in imaging applications to selectively label F-actin. Fluorescently-labeled phalloidin has several advantages over antibodies for actin labeling, including virtually identical binding properties with actin from different species of plants and animals, and low non-specific binding. Phalloidin binds F-actin with high selectivity, while Alexa Fluor Plus 405 provides violet fluorescence of unparalleled brightness and photostability.

Use in multiple applications
Alexa Fluor Plus 405 Phalloidin can be used to visualize and quantitate F-actin in tissue sections, cell cultures, or cell-free preparations. Its staining is fully compatible with other fluorescent stains used in cellular analyses, including fluorescent proteins, Qdot nanocrystals, and other Alexa Fluor conjugates including secondary antibodies.

Pierce™ Protein G Magnetic Beads Thermo Scientific™

Thermo Scientific Pierce Protein G Magnetic Beads are high-capacity and high-throughput affinity particles for antibody purification and immunoprecipitation methods using manual or robotic magnetic separators.

Features of Protein G Magnetic Beads:

High efficiency – equivalent or higher yield of IP target antigens than magnetic beads from other suppliers
Low non-specific binding – stable, pre-blocked beads provide highly purified product (e.g., antigen eluted in IP with antibody is devoid of contaminating protein from complex cell lysate)
Consistent – magnetic beads eliminate resin loss and provide for more efficient separation of solutions than traditional IP methods that use only centrifugation
Versatile – beads are compatible with manual and automated workflows (e.g., Thermo Scientific KingFisher Instruments)

Recombinant protein G, covalently attached to a blocked magnetic bead surface, can bind to antibodies from many different species, enabling purification of antibodies from crude extracts. Immunoprecipitation assays performed with protein G coated beads result in high yield of target antigen with very low background. Using our crosslinker chemistry, you can immobilize an antibody onto the magnetic particle and prevent IgG contamination in IP or Co-IP experiments. The Pierce Protein G Magnetic Beads can be used both manually with a magnetic stand as well as with automated platforms such as the Thermo Scientific KingFisher Instruments for high-throughput workflows.

Applications:
• IP and Co-IP experiments
• Antibody purification

Thermo Scientific Pierce Protein G Magnetic Beads are used for purifying antibody from serum, cell culture supernatant or ascites as well as for IP/Co-IP of antigens from cell or tissue extracts. Protein G can bind to antibodies from many different species, including mouse, human, rabbit, cow, goat and sheep. The protocol for the Pierce Protein G beads has been optimized to allow for high recovery and high purity of the isolated antibody or antigen. Antibody or antigen/antibody complex (IP) is first captured on the magnetic beads. The beads are washed and then the target is eluted with low pH elution buffer. IP performance is equivalent to or better than Protein G magnetic beads from other suppliers.

Pierce™ Crosslink Magnetic IP/Co-IP Kit Thermo Scientific™

The Thermo Scientific Pierce Crosslink Magnetic IP/Co-IP Kit uses crosslinking chemistry to covalently immobilize IP antibodies onto premium-quality Protein A/G Magnetic Beads for effective immunoprecipitation and co-immunoprecipitation.

Because this magnetic IP procedure involves covalent attachment of the specific antibody used for immunoprecipitation, target proteins or co-IP protein complexes can be eluted and analyzed without antibody fragments because the antibody remains affixed to the beads. The kit contains an optimized protocol and all buffers and reagents necessary to accomplish high-yield IP or co-IP with antibodies that bind to Protein A/G and using either manual or automated magnetic separation tools.

Features of the Crosslink Magnetic IP/Co-IP Kit:

Antibody-free IP—antibody is irreversibly attached to the magnetic beads resulting in negligible co-elution of intact antibody or heavy and light chains with the purified antigen
Convenient—the IP/co-IP kit contains magnetic beads and all essential buffers for optimal immunoprecipitation
Fast—complete crosslinking and immunoprecipitation in less than 3 hours
Efficient—immunoprecipitate with half the recommended volume of magnetic particles compared to other magnetic beads
Compatible—magnetic beads coupled with Protein A/G recombinant protein ensures compatibility with most primary antibodies, whether from mouse or rabbit
Scalable—use only the amount of antibody needed for a single IP experiment or prepare a larger quantity of antibody-crosslinked magnetic beads for multiple experiments

The protocol for the Pierce Crosslink Magnetic IP/Co-IP Kit binds IP antibody to Protein A/G Magnetic Beads and then covalently crosslinks the antibody to the beads using disuccinmidyl suberate (DSS). The antibody-crosslinked beads are then incubated with cell lysate or tissue extract that contains the protein antigen of interest, allowing the antigen:antibody complex to form. The beads are washed to remove non-bound material and a low pH elution buffer is used to dissociate bound antigen from the antibody-crosslinked beads. Neutralization buffer is included to prevent precipitation of the isolated antigen and to ensure protein activity in downstream applications. Lane Marker Sample Buffer is included for preparing samples for SDS-PAGE analysis. The protocol is optimized for 2 to 10µg of IP antibody. For optimal co-IP yields, use 5 to 10µg of antibody. The beads are removed from the solution manually using a magnetic stand or by automation with an instrument such as the Thermo Scientific KingFisher Flex Instrument.

Traditional IP (without covalent antibody attachment) generally provides higher antigen yields than IP protocols that used crosslinking. This is because the crosslinking process inevitably causes loss of some functional antibody binding sites. To overcome this limitation with the crosslink method, it may be necessary to use somewhat greater amounts of IP antibody in the crosslink IP method than in the equivalent traditional IP procedure. Of course, the advantage with the crosslink immunoprecipitation procedure is Western blots devoid of interfering antibody bands.

Protocol Summary
1: Prewash beads with 1X Coupling Buffer.
2: Bind antibody to Protein A/G magnetic beads for 15 minutes.
3: Wash beads three times with 1X Coupling Buffer.
4: Crosslink antibody to beads with DSS for 30 minutes.
5: Wash bead three times with Elution Buffer and then twice with IP Lysis/Wash Buffer.
6: Incubate cell lysate with prepared beads for 1-2 hours at room temperature or overnight at 4°C.
7: Wash beads twice with IP Lysis/Wash Buffer and then once with purified water.
8: Elute bound antigen.

Related Products
Pierce™ Crosslink IP Kit

Click-iT™ Alexa Fluor™ 488 sDIBO Alkyne for Antibody Labeling Invitrogen™

The Click-iT Alexa Fluor 488 sDIBO Alkyne for Antibody Labeling is optimized for easy attachment to azido modified antibodies using copper-free Click chemistry. This sDIBO label can be used with antibodies that have been modified using the SiteClick Antibody Azido Modification Kit or antibodies that have been engineered to contain azido moieties. These sDIBO alkynes are improved versions of our original DIBO cyclooctynes, yielding conjugates that are less 'sticky' and give lower signal background in biological samples.

This modular labeling system gives you the option to choose different fluorescent labels for your antibody and attach another molecule via streptavadin or your own molecule via amine-reactive or amine-containing moieties depending on your assay.

There are multiple Click-iT sDIBO labels to choose from:
Click-iT Alexa Fluor 488 sDIBO Alkyne for Antibody Labeling
Click-iT Alexa Fluor 555 sDIBO Alkyne for Antibody Labeling
Click-iT Alexa Fluor 647 sDIBO Alkyne for Antibody Labeling
Click-iT Biotin sDIBO Alkyne for Antibody Labeling
Click-iT Amine sDIBO Alkyne for Antibody Labeling
Click-iT SDP Ester sDIBO Alkyne for Antibody Labeling

Learn more about SiteClick labeling technology ›

Custom SiteClick Antibody Labeling Service and sDIBO labels
If you have an antibody that is considered 'difficult to label' or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

LanthaScreen™ Thiol Reactive Tb Chelate Thermo Scientific™

As part of the LanthaScreen® TR-FRET toolbox of assay reagents, LanthaScreen® Thiol Reactive Tb Chelate labeling reagent is available for assay development. The thiol-reactive maleimide group can be conjugated to virtually any peptide or protein that contains one or more accessible thiol moieties, such as the thiol group found on a cysteine residue. Energy transfer from the terbium donor to a suitable acceptor such as fluorescein is readily detected by monitoring an increase in acceptor fluorescence intensity. See the user guide for an application-based protocol for cysteine-containing peptides or proteins.

MultiShot™ TOP10 Chemically Competent E. coli Invitrogen™

MultiShot™ TOP10 Chemically Competent E. coli provide transformation efficiencies of >1 x 108 cfu/µg control plasmid DNA, and are suited for high-throughput transformations. MultiShot™ TOP10 Chemically Competent cells are packaged in five 96-well microtiter plates (15 µL aliquots) to simplify high-throughput bacterial transformations. Benefits of MultiShot™ TOP10 cells:

• Designed for high-throughput applications—microtiter plate format designed for automated high-throughput cloning
• Compatible with genomic DNA—the mcrA mutation supports efficient transfection of methylated DNA
• Permits rapid screening—the LacZΔM15 allele permits rapid blue/white screening of transfectants on plates containing X-Gal or Bluo-Gal
• Efficient—the endA1 mutation helps reduce endonuclease activity, improving subsequent plasmid DNA yields
• Reliable—the recA1 mutation helps reduce unwanted recombinationEasy to use for high-throughput transformations
MultiShot™ TOP10 Chemically Competent cells are genetically similar to the reliable DH10B™ strain, and are available in five 96-well plates to facilitate high-throughput transformation. After addition of DNA, MultiShot™ TOP10 Chemically Competent cells can be transformed by heat-shock at 42°C using a heat block or thermocycler. Using our TA Cloning® and TOPO® Cloning protocols, including controls, yields of 100–400 colonies per agar plate can be expected.

Genotype: F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara, leu)7697 galU galK rpsL (StrR) endA1 nupG

Find the strain and format that you need
We also offer many other different strains and formats of chemically competent cells and electrocompetent cells to help meet your specific transformation needs. If you require other high-throughput transformation options, choose from our collection of MultiShot™ formatted comp cells or contact us for custom formatting options.

Pierce™ Streptavidin Magnetic Beads Thermo Scientific™

Thermo Scientific Pierce Streptavidin Magnetic Beads accelerate throughput for automated magnetic purification of biotinylated molecules.

Features of Streptavidin Magnetic Beads:

High-performance beads—non-aggregating, pre-blocked, iron oxide, superparamagnetic microparticles provide exceptional uniformity for automated HTS and manual applications alike
Stable immobilization chemistry—streptavidin is immobilized using leach-resistant chemistry
High capacity—superior quality beads with high binding capacity provide rapid and efficient biomolecule purification from complex samples
Low non-specific binding—stable, pre-blocked beads provide clean purification products (e.g., antigen eluted in IP with biotinylated antibody) that are compatible with mass spectrometry analysis and other downstream analyses
Superior performance—nearly three times higher binding capacity than typical beads from other suppliers, allowing the use of smaller amounts per experiment

Applications:
• Immunoprecipitate antigens (using biotinylated antibodies) from a wide variety of sources
• Co-immunoprecipitate interaction complexes using biotinylated antibodies
• Capture protein-protein interactions in pull-down assays using biotinylated 'bait' proteins
• Isolate biotin-labeled DNA-protein complexes from cell or tissue extracts
• Capture single-stranded biotinylated DNA oligos
• Isolate biotinylated PCR products

These streptavidin magnetic beads are validated and optimized for use with high-throughput magnetic platforms, such as the Thermo Scientific KingFisher 96 and KingFisher Flex Instruments, but the beads also enable premium performance for simple benchtop applications using an appropriate magnetic stand. The iron oxide, super-paramagnetic particles offer superior performance (high capacity and low nonspecific binding) compared with other commercial magnetic beads.

Pierce Streptavidin Magnetic Beads use a recombinant form of streptavidin with a mass of 53kDa and a near-neutral isoelectric point (pI). The protein is a tetramer having four biotin-binding sites. Unlike avidin, streptavidin has no carbohydrate groups, resulting in low nonspecific binding. The high-affinity interaction between streptavidin and biotin cannot be dissociated efficiently except with very harsh conditions, such as boiling in sample loading buffer for SDS-PAGE or 8M guanidine·HCl, pH 1.5. Consequently, it is often possible to elute binding partners in an interaction complex without also eluting the biotinylated component.

Phalloidin Invitrogen™

Unlabeled phalloidin can be used as a control in blocking F-acting staining or in promoting actin polymerization. can be used as a control in blocking F-acting staining or in promoting actin polymerization.

Pierce™ Protein L Magnetic Beads Thermo Scientific™

Thermo Scientific Pierce Protein L Magnetic Beads are high-capacity and high-throughput affinity particles for antibody purification using manual or robotic magnetic separators.

Features of Protein L Magnetic Beads:

Selective—immobilized Protein L is ideal for selective purification of human and mouse antibodies that have kappa light chains
Low non-specific binding—stable, pre-blocked beads provide clean purification of antibody
Versatile—beads are compatible with manual and automated workflows (e.g., Thermo Scientific KingFisher Instruments)

Recombinant Protein L, covalently attached to a blocked magnetic bead surface, selectively binds mouse and human antibodies through kappa light chains. The beads are commonly used to purify monoclonal antibodies in cell culture supernatants supplemented with bovine serum as Protein L does not bind bovine IgG. The Pierce Protein L Magnetic Beads can be used manually with a magnetic stand, as well as with automated platforms such as the Thermo Scientific KingFisher Instruments for high-throughput workflows.

Applications:
• Purification of monoclonal and polyclonal antibodies that bind poorly to Protein A or Protein G magnetic beads
• Purification of monoclonal antibodies from culture supernatants supplemented with bovine serum
• Protein L IP and co-IP assays
• Purification of ScFv and Fab fragments containing kappa light chains

Thermo Scientific Pierce Protein L Magnetic Beads are typically used for purifying mouse and human antibodies containing kappa light chains from serum, cell culture supernatant or ascites. Protein L can bind a broader range of Ig classes than Protein A or Protein G, including IgG, IgM, IgA, IgE and IgD. Protein L binds strongly to human (kappa I, III and IV only), mouse (kappa I only), rat and pig immunoglobulins. It binds weakly to rabbit immunoglobulins and does not bind bovine, goat or sheep immunoglobulins. Single chain variable fragments (scFv) and Fab fragments also bind to Protein L. The protocol for the Pierce Protein L beads has been optimized to allow for high recovery and high purity of isolated antibodies.

Click-iT™ Alexa Fluor™ 555 sDIBO Alkyne for Antibody Labeling Invitrogen™

The Click-iT Alexa Fluor 555 sDIBO Alkyne for Antibody Labeling is optimized for easy attachment to azido modified antibodies using copper-free Click chemistry. This sDIBO label can be used with antibodies that have been modified using the SiteClick Antibody Azido Modification Kit or antibodies that have been engineered to contain azido moieties. These sDIBO alkynes are improved versions of our original DIBO cyclooctynes, yielding conjugates that are less 'sticky' and give lower signal background in biological samples.

This modular labeling system gives you the option to choose different fluorescent labels for your antibody and attach another molecule via streptavadin or your own molecule via amine-reactive or amine-containing moieties depending on your assay.

There are multiple Click-iT sDIBO labels to choose from:
Click-iT Alexa Fluor 488 sDIBO Alkyne for Antibody Labeling
Click-iT Alexa Fluor 555 sDIBO Alkyne for Antibody Labeling
Click-iT Alexa Fluor 647 sDIBO Alkyne for Antibody Labeling
Click-iT Biotin sDIBO Alkyne for Antibody Labeling
Click-iT Amine sDIBO Alkyne for Antibody Labeling
Click-iT SDP Ester sDIBO Alkyne for Antibody Labeling

Learn more about SiteClick labeling technology ›

Custom SiteClick Antibody Labeling Service and sDIBO labels
If you have an antibody that is considered 'difficult to label' or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

DyLight™ 488 NHS Ester Thermo Scientific™

Thermo Scientific DyLight 488 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 488 for fluorescent labeling of antibodies and other proteins to be used as molecular probes for cellular imaging and other fluorescence detection methods.

DyLight 488 has high fluorescence intensity over a broad pH range (pH 4-9) and is more photostable than Cy2™, Alexa™ Fluor 488, FITC and LI-COR™ dyes in many applications. The high water solubility of DyLight Fluors allows a high dye-to-protein ratio to be achieved without causing precipitation of the conjugates. DyLight 488 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.

Features of the DyLight 488 NHS Ester:

High performance— DyLight 488 is comparable to Alexa Fluor 488 and brighter than FITC and Cy2
Specific— NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
Optimized procedure— following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling

Applications:
• Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
• Target protein labeling for in vitro and in vivo fluorescent detection strategies

DyLight 488 Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. Learn more about NHS ester chemistry.

Typical labeling reactions require the dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to specific amounts of the labeling reagent. This feature allows DyLight 488 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

We also offer Standard and Microscale DyLight 488 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods.The standard size kit contains all necessary components to perform three separate labeling reactions using 1 mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG. Both kit sizes include the Amine-Reactive DyLight 488 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.

Related Products
DyLight™ 488 Antibody Labeling Kit
DyLight™ 488 Microscale Antibody Labeling Kit

ProteinSEQ™ HCP Core Kit Applied Biosystems™

The ProteinSEQ™ HCP Core Kit is a customizable kit that enables users to customize their ProteinSEQ host cell protein measurements for their own cell lines. It contains buffers and reagents needed to perform 200 PCR reactions to quantify host cell proteins of any cell line for which there is an antibody available.

Features include:

• Precise—quantitate host cell protein contaminants with excellent precision
• Fast—helps accelerate process development with more valid test results per run
• Efficient—helps reduce hands-on time with less sample dilution and preparation
• Reproducible—achieve unprecedented productivity with 4-log dynamic range

ProteinSEQ CHO HCP quantitation
The ProteinSEQ assay platform uses qPCR technology to measure protein process contaminants with extraordinary sensitivity and unparalleled dynamic range. Host cell protein contaminants from any cell line can be measured easily and effectively by incorporating any anti-HCP antibody into the ProteinSEQ workflow.

Streamlined workflow
The ProteinSEQ HCP Core Kit is the only instrument platform-based HCP quantitation solution to deliver a customizable HCP assay with class-leading sensitivity and 4-log dynamic range. The kit helps accelerate laboratory workflows because the number of sample dilutions and replicates is reduced in each run.

Sulfo-SBED Biotin Label Transfer Kit - Western Blot Application Thermo Scientific™

The Thermo Scientific Pierce Sulfo-SBED Biotin Label Transfer Reagent is a multifunctional reagent for labeling a purified protein and then covalently transferring the attached biotin tag onto specific interactors of that protein.

Sulfo-SBED is the abbreviation for Sulfo-N-hydroxysuccinimidyl-2-(6-[biotinamido]-2-(p-azido benzamido)-hexanoamido) ethyl-1,3'-dithioproprionate. It is a heterobifunctional chemical crosslinker capable of covalently attaching to primary amines at one end and to nearly any protein functional group at the other end. Unlike typical crosslinkers, Sulfo-SBED also includes a biotin group and a cleavable disulfide spacer arm. Together these features allow one to sequentially crosslink interacting proteins and transfer the biotin affinity tag from one protein (i.e., a purified 'bait' protein) to another (possibly unknown 'prey' protein). Label Transfer is a powerful in vitro method for protein interaction discovery. A growing number of publications feature the use of Sulfo-SBED Biotin Label Transfer Reagent to identify previously unknown protein interaction binding partners and to more fully characterize the specific protein binding domains of other protein interactions.

Typical protocol for label transfer experiment:

• Add a few microliters of dissolved Sulfo-SBED Reagent to 0.5-1 mL of purified bait protein in PBS.
• Incubate mixture for 30-120 minutes on ice or at room temperature in the dark.
• Desalt or dialyze (in subdued light) to remove excess non-reacted Sulfo-SBED from the labeled bait protein.
• Add labeled bait protein to cell lysate or other solution containing putative target protein interactors ('prey').
• When interaction complexes have formed, expose the solution to ultraviolet light (365 nm) for several minutes.
• Analyze products by one of several methods:
Western Blotting: Cleave crosslinks in DTT, separate proteins by SDS-PAGE, and detect biotinylated bands by Western blotting with streptavidin-HRP.
Purification and Mass Spec or Sequencing: Affinity-purify biotinylated proteins or peptide fragments following trypsin digestion and perform MS or sequencing to characterize the proteins involved.

Related Products
Sulfo-SBED Biotin Label Transfer Reagent

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