Pierce™ Antibody Biotinylation Kit for IP provides biotinylation reagents designed specifically for the labelling of primary antibodies used in immunoprecipitation applications.

• Easy to use, kit configuration specifically optimised to accommodate 50 to 200 μg of antibody in a 100 μl reaction
• Zeba™ desalting columns are provided to remove excess biotinylation reagent after labelling
• Reacts with primary amines (-NH₂) of antibodies
• Pegylation improves water solubility of the biotinylated antibody and prevents aggregation upon storage
• Longer spacer arm (29 angstroms) minimize steric hindrance when binding to avidin molecules

These reagents have been optimised 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 labelling procedure in this kit for IP has been developed to address this challenge.

The NHS-PEG4-biotin labelling reagent contains an amine-reactive N-hydroxysuccinimide ester (NHS) group and a water-soluble PEG4 spacer for optimal labelling. Both the labelling efficiency of the biotinylation reagent and binding affinity of the labelled antibody have been validated using mouse monoclonal (IgG1, IgG2), rabbit polyclonal, and monoclonal antibodies.