Pierce™ HA-Tag IP/Co-IP Kit (Thermo Scientific™)

The Thermo Scientific Pierce HA Tag IP/Co-IP Kit provides the affinity resin, positive control and other reagents necessary to perform immunoprecipitation (IP) or co-immunoprecipitation (Co-IP) reactions with an HA-tagged bait protein.

Features of the HA-Tag IP/Co-IP Kit:

• Improved—updated kit includes more immobilized antibody resin and improved protocol along with GST-PI3K-SH2-HA fusion protein as a positive control
• Specific—highly specific anti-HA monoclonal antibody provides high-yield immunoprecipitation products and clean Western blot detection
• Compatible—includes protocols and reagents for multiple elution conditions to accommodate different protein sensitivities and downstream applications
• Robust—the kit and the anti-HA agarose are compatible with various cell lysates and physiologic (non-denaturing) buffer systems
• Convenient and easy—complete kit includes all necessary reagents, convenient spin columns and easy-to-follow instructions

This kit is based on crosslinked beaded agarose to which a highly specific anti-HA antibody is covalently immobilized. The ready-to-use affinity resin, together with the included buffers, microcentrifuge spin columns, positive control, and easy-to-follow instructions, constitute a complete set of reagents sufficient for 25 for IP or Co-IP assays. Upon incubation with a sample containing the tagged fusion protein, interaction complexes involving the HA-tagged bait protein are captured on the agarose beads. After simple washing steps, the specific protein interaction complex is easily eluted from the resin in the supplied elution buffer or SDS-PAGE sample loading buffer for subsequent analysis.

The hemagglutinin (HA) peptide (YPYDVPDYA), derived from the human influenza virus HA protein is one of several fusion protein tags used for recombinant protein expression. Utilizing a specific, high-affinity immobilized antibody, HA-tagged fusion proteins can be quickly purified from bacterial and mammalian cell lysates as well as from the Pierce Human in vitro translation reactions. For co-immunoprecipitation reactions, simple wash steps allow enrichment and elution of specific protein interaction complexes into the supplied elution buffer or SDS-PAGE sample loading buffer for subsequent analysis.

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