Next‐generation Immunoprecipitation Kits Validated for Functional Applications

Abstract

The study of protein function and interaction is essential for understanding many biological processes. We have developed three new immunoprecipitation (IP) kits with improved protocols and buffers that provide excellent antigen recovery with the least amount of nonspecific binding using 2 to 10 µg of antibody. The Pierce IP Direct kit contains our AminoLink Plus Coupling resin for directly crosslinking purified antibodies to the matrix for specific elution of the antigen. The Pierce IP Classic kit contains Protein A/G Plus Agarose that binds immunoglobulins from a variety of species. The Protein A/G Plus resin is also used in the Pierce IP Crosslinked kit which is designed to purify and crosslink the antibody to the resin before performing the immunoprecipitation procedure. With the new kits, we have demonstrated immunoprecipitations of a diverse selection of proteins including membrane proteins, protein phosphatases and kinases, transcription factors, and cell cycle regulation proteins. We show that the new Thermo Scientific Pierce IP Kits immunoprecipitate protein complexes and are compatible with mass spectrometry and enzymatic assays.