The Thermo Scientific Pierce Classic IP Kit uses high-quality Protein A/G agarose beads, microcentrifuge spin columns and buffers to perform traditional immunoprecipitation assays 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.
- Assay consistency – 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 – purify 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
- Routine, traditional immunoprecipitation experiments
- Immunoprecipitation for analysis in nonreducing conditions
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.
- Petersen, H.H., et al. (2003). J. Cell Sci. 116, 453-461.
Review of immunoprecipitation techniques and methods
Pierce IP Kit Selection Guide
Other immunoprecipitation kits – agarose, magnetic, co-IP, etc.
Control Agarose Resin – for pre-clearing samples and various negative controls for IP
Spin Columns - Screw Cap – the same spin columns (Part No. 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