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Thermo Scientific™ Pierce™ Co-Immunoprecipitation Kit

Catalog No. PI26149
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50 reactions kit
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PI26149 50 reactions kit
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Catalog No. PI26149 Supplier Thermo Scientific™ Supplier No. 26149
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Description

Covalently immobilize IP antibodies and perform co-IP assays in which target protein complexes are immunoprecipitated without antibody contamination.

Thermo Scientific™ Pierce Co-Immunoprecipitation Kit provides all the components necessary to perform a properly controlled co-IP experiment and obtain purified products without antibody contamination.

The Co-Immunoprecipitation (Co-IP) Kit enables isolation of native protein complexes from a lysate or other complex mixture by directly immobilizing purified antibodies onto an agarose support. Co-IP is a common approach to study protein:protein interactions that uses an antibody to immunoprecipitate the antigen (bait protein) and co-immunoprecipitate any interacting proteins (prey proteins). Traditional co-IP methods that use Protein A or G result in co-elution of the antibody heavy and light chains that may co-migrate with relevant bands, masking important results. The Pierce Co-IP Kit resolves this issue by covalently coupling antibodies onto an amine-reactive resin. The kit includes optimized buffers for protein binding and recovery, reagents to perform control experiments and efficient spin columns and collection tubes, which shorten the protocol and minimize handling and mixing.

Highlights:

  • Works for any purified antibody – perform co-immunoprecipitation experiments using any purified antibody regardless of species or Ig class (chicken IgY, human IgE, mouse IgG1, IgM, etc.); no dependence on Protein A and Protein G affinity binding

  • No antibody interference – covalent attachment method and non-reducing elution system yields pure co-IP products without contamination by the IP antibody

  • Reusable antibody beads – covalent antibody immobilization allows prepared IP resin to be reused and provides potential savings of costly antibody

  • Complete Kit – all reagents for antibody immobilization and many IP experiments, including binding and wash buffers, elution buffer and convenient microcentrifuge spin columns to make resin manipulations easier and more efficient

  • Control beads included – underivatized agarose beads provided for use as negative control for nonspecific binding

  • Versatile – co-IP method is compatible with any physiolgical (non-denaturing) protein sample buffer that is compatible with the specific antibody-antigen and protein interaction complex

Includes:

AminoLink Plus Coupling Resin, coupling buffer, quenching buffer, wash solution, sodium cyanoborohydride solution, IP lysis/wash buffer, conditioning buffer, Tris-buffered saline, elution buffer, lane marker sample buffer, control agarose resin, spin columns and collection tubes

Specifications

Content And Storage Sufficient For: Preparing and performing 50 co-IP reactions, each with 25 μL of resin
• AminoLink Plus Coupling Resin, 2 mL
• Coupling Buffer (20X), 25 mL
• Sodium Cyanoborohydride Solution (5M), 0.5 mL
• Quenching Buffer, 50 mL
• Wash Solution, 60 mL
• IP Lysis/Wash Buffer, 2 x 50 mL
• Modified Dulbecco's PBS (20X), 25 mL
• Conditioning Buffer (100X), 5 mL
• Elution Buffer, 50 mL
• Lane Marker Sample Buffer (5X), 5 mL
• Pierce Spin Columns, Screw Cap, 50 columns
• Microcentrifuge Collection Tubes, 2 mL, 100 tubes
• Microcentrifuge Collection Tubes, 1.5 mL, 50 tubes
• Pierce Control Agarose Resin, 2 mL

Store at 4°C.
Assay Affinity Purification
For Use With (Equipment) Microcentrifuge
Product Line Pierce
Quantity 50 reactions kit
For Use With (Application) Immunoprecipitation (IP)
Target Not Target-Specific
Type Immunoprecipitation/Co-Immunoprecipitation Kit
Sufficient For 50 Reactions
Format Kit

Frequently Asked Questions (FAQs)

What is the shelf life of the Pierce Co-Immunoprecipitation Kit (Cat. No. 26149)?

The Pierce Co-Immunoprecipitation Kit is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended (4 degrees C). Please see section 8.1 of our Terms & Conditions of Sale for more details.
Can I use samples harvested using my own Lysis Buffer with the Thermo Scientific Co-Immunoprecipitation Kit?

Yes, but the IP Lysis/Wash Buffer is formulated to be an efficient lysing agent that is gentle to proteins. The exclusion of ionic detergents and pH extremes help to preserve protein: protein interactions improving the results of recovering both antigen and its interacting partners at the elution step (versus interating partners being lost in the wash steps).
Why does the Thermo Scientific Co-Immunoprecipitation Kit include a Conditioning Buffer (100X)?

The conditioning buffer helps with the wash and especially to provide the correct environment to enhance the efficiency of the low pH Elution Buffer. The overall result is more antigen recoved in the first volume of elution (more concentrated antigen).
Can I choose to immobilize an un-purified antibody with the Thermo Scientific Co-Immunoprecipitation Kit?

No, we do not recommend immobilizing an un-purified antibody; a purified antibody is required. The AminoLink Coupling Resin that is included will react with primary amines that are found on the surface of virtually all proteins, resulting in immobilization of many other proteins along with the antibody. This will reduce the amount of antibody immobilized, resulting in less antigen and interacting partners being purified. It can also result in higher background as other proteins in the sample may bind to the other immobilized proteins and co-elute with the antigen and interacting partners, contaminating them.

The protocol recommends using 10 - 75 µg of antibody with the Thermo Scientific Co-Immunoprecipitation Kit, is it possible to use a different amount?

Yes, the product manual includes “Table 1. Amounts of antibody, coupling resin, wash buffer and elution buffer to use” that allows researchers to scale their affinity purification using 5 - 1,000 µg of antibody. We do not recommend going below 5 µg of antibody. This was the smallest amount of antibody tested inhouse. The rationale to stop at 5 µg of antibody is that it was felt that an extremely strong affinity between the antibody and antigen would be required to get sensitive, reproducible results with less antibody to capture the antigen and interacting partners.

For Research Use Only. Not for use in diagnostic procedures.