Thermo Scientific Pierce Cross-Linked IP Kits use traditional IP methods to include cross-linking of IP antibodies to Protein A/G agar Sugar reagents and protocols to achieve antigen immunoprecipitation without antibody contamination.

The main benefits resulting from these features are the ability to purify target proteins without antibody contamination and the ability to more efficiently wash and separate samples from beaded agarose resin.

Features of the Cross-Linked IP Kit:

•Eliminates Antibody Contamination—Irreversibly link antibodies to agarose beads, minimizing heavy chain and Light chains co-elute with purified proteins
•Simple and efficient scalability—Prepare off-the-shelf IP affinity using only the amount of antibody required for a single IP experiment or immobilize 100-200 μg of antibody Resins are used in many experiments
•Assay reliability and sample handling—Pierce spin columns eliminate resin loss and provide solution separations more efficiently than traditional IP methods using only microcentrifuge tubes
•Prepared antibody affinity resins are reusable—antibodies are covalently immobilized and are typically not inactivated by mild elution procedures, so the resin can often be reused several times
• Complete Kit—Package includes sufficient reagents and spin columns for at least 50 antibody immobilization and IP experiments

Applications:
• Subsequently passed Immunoprecipitation and SDS-PAGE analysis of target proteins with molecular weights similar to heavy or light chain antibody fragments
i.e., methods that do not involve SDS-PAGE) immunoprecipitation of the target protein for subsequent analysis

The Pierce Cross-Linked IP Kit method involves capturing the IP antibody onto Protein A/G Sepharose resin and passing it through the Disuccinimide diacid (DSS) cross-linking covalently fixes it to the support. The antibody resin is then incubated with a sample containing the target protein antigen, allowing antibody:antigen complexes to form. After washing to remove non-bound (potentially unwanted) components from the sample, the antigen is recovered by dissociating it from the antibody using the elution buffer provided in the kit. Complete the entire procedure in a microcentrifuge cup so that the solution is completely separated from the agarose resin after a brief centrifugation. Only if the antigen is eluted by this procedure can the antigen be identified and further analyzed without interference from antibody fragments. Additionally, the antibody resin can often be reused for additional rounds of immunoprecipitation.

Related products
Pierce™ cross-linked magnetic bead method IP /Co-IP Kit