A convenient kit for a new spin on traditional immunoprecipitation.

The Thermo Scientific Pierce Classic IP Kit provides all the necessary reagents, spin cups and collection tubes to perform successful immunoprecipitation (IP) experiments with ease. The kit uses high-capacity Protein A/G agarose affinity resin for efficient binding of most species and subclasses of IP antibodies. The included IgG elution buffer provides milder and less denaturing recovery of antibody:antigen complexes than the traditional method of boiling in reducing sample buffer for SDS-PAGE, facilitating a greater variety of methods for subsequent analysis. The microcentrifuge spin column format helps to ensure effective washing and separation of samples from the beaded agarose affinity resin.

Like traditional IP methods, the Pierce Classic IP Kit procedure involves formation of antibody:antigen complexes in a sample solution and then capture of that complex to an IgG-binding protein that is covalently bound to beaded agarose resin (Protein A/G Agarose). After washing to remove nonbound (presumably undesired) components of the sample, the antigen and antibody are recovered from the beaded resin with elution buffer supplied in the kit. The entire procedure is performed in a microcentrifuge spin column, allowing solutions to be fully separated from the agarose resin upon brief centrifugation.

The Thermo Scientific Pierce Classic IP Kit uses high-capacity Protein A/G Agarose to capture and purify target antigen through its specific antibody.

Highlights:

• Assay consistency – Pierce Spin Columns (see dimensions) eliminate resin loss and provide for more efficient separation of solutions than traditional IP methods that use only microcentrifuge tubes
• Fast – Immunoprecipitate (IP) in less than one hour
• Mild elution conditions – Recover antigen without harsh detergents or reducing agents
• Easy-to-follow instructions – IP target protein from crude lysate in four simple steps
• Suitable for most common antibody types – Protein A/G provides excellent binding for most mouse, rabbit, human and goat IgG subclasses
• Complete kit – includes lysis/wash buffer, binding and elution buffers, control agarose resin, sample loading buffer and spin columns

Applications:

• Routine, traditional immunoprecipitation experiments
• Immunoprecipitation for analysis in nonreducing conditions

How can I be sure that the Protein A/G Agarose will bind to my antibody?
Protein A/G is a recombinant protein that combines antibody-binding domains from Protein A and Protein G. Typically, Protein A is preferred for use with Rabbit polyclonal antibodies, while Protein G is preferred for use with mouse antibodies (especially monoclonals of the IgG1 subclass).
Compare Protein A, Protein G and other antibody-binding proteins
Look up Protein A/G binding characteristics by antibody species and class

Which of the three different kinds of Pierce IP Kits is best for me?
Compare the features and requirements of Thermo Scientific Pierce Classic, Crosslink and Direct IP Kits to determine which one is most appropriate for an intended IP experiment and available source of antibody.

Related Products:

Crosslink IP Kit – for antibody immobilization by crosslinking
Direct IP Kit – for immunoprecipitation using any species of purified antibody
Co-IP Kit – for co-immunoprecipitation using the Direct IP method

Control Agarose Resin – for pre-clearing samples and various negative controls for IP
Spin Columns - Screw Cap – the same spin columns (#69705) supplied in the kit
Pierce IP Lysis Buffer – best cell lysis buffer for immunoprecipitation
M-PER Mammalian Protein Extraction Reagent – alternative, IP-compatible cell lysis buffer
Protein A/G Agarose and UltraLink Resins – for antibody purification

References:

1. Petersen, H.H., et al. (2003). J. Cell Sci. 116, 453-461.