The Thermo Scientific™ Pierce™ Antibody Biotinylation Kit for IP provides biotinylation reagents designed specifically for the labeling of primary antibodies used in immunoprecipitation applications.Optimized for IP—reagents and protocols developed specifically to address antibody labeling for immunoprecipitation applicationsEasy to use—kit configuration specifically optimized to accommodate 50–200 µg of antibody in a 100 µL reaction; desalting columns are provided to remove excess biotinylation reagent after labelingAmine reactive—reacts with primary amines (-NH2) of antibodiesEnhanced solubility—pegylation improves water solubility of the biotinylated antibody and prevents aggregation upon storageReduced steric hindrance—longer spacer arm (29 angstroms) minimize steric hindrance when binding to avidin moleculesThe Pierce Antibody Biotinylation Kit reagents have been optimized and validated to biotin label antibodies for IP and co-IP reactions

Determining the optimal number of biotins to attach to the target molecule is one of the major challenges of biotinylation. For IP and co-IP applications, too many biotins result in reduced affinity for the target antigen, while too few biotins result in antibody leaching upon elution of the target antigen. The biotin labeling procedure in the Pierce Antibody Biotinylation Kit for IP has been developed to address this challenge.The kit contains sufficient reagents to label 50–200 µg of antibody in 100 µL reaction volumes for eight samples. The NHS-PEG4-biotin labeling reagent contains an amine-reactive N-hydroxysuccinimide ester (NHS) group and a water-soluble PEG4 spacer for optimal labeling and is provided in easy-to-use, single-use microtubes. Both the labeling efficiency of the biotinylation reagent and binding affinity of the labeled antibody have been validated using mouse monoclonal (IgG1, IgG2), rabbit polyconal, and rabbit monoclonal antibodies. Zeba™ Desalting Spin Columns are provided for easy and efficient removal of salts and excess biotin.Applications:IP and co-IPIP and co-IP for mass spectrometry applicationsAntibody attachment to streptavidin agarose and magnetic supports