The Thermo Scientific Pierce Magnetic HA-Tag IP/Co-IP Kit contains specific immunoaffinity magnetic beads and reagents to perform immunoprecipitation assays of HA fusion proteins or co-IP experiments using HA-tagged bait proteins.

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

• Specific magnetic beads—covalently immobilized high-quality anti-HA monoclonal antibody enables high yields of immunoprecipitation products
• Low non-specific binding—stable, pre-blocked beads and specific antibody minimize off-target binding
• Trouble-free elution—low-pH elution buffer ensures recovery of HA-tagged protein interaction complexes without antibody leaching contamination
• Convenient and fast—complete kit and easy-to-follow instructions provide optimized protocols to perform IP or Co-IP experiment in approximately 1 hour
• Versatile—magnetic beads are compatible with manual and automated magnetic separation workflows (e.g., Thermo Scientific KingFisher Instruments)

Unlike traditional IP procedures based on capture with Protein A/G beads, this kit uses magnetic beads containing pre-immobilized anti-HA antibody. These Pierce Anti-HA Magnetic Beads ensure specific binding of HA-tagged protein complexes from biological samples. Because the antibody is covalently attached to the beads, IP-targets are easily eluted and recovered without antibody contamination. The complete IP kit includes the magnetic beads, lysis/wash buffer, low-pH elution buffer, neutralization buffer, HA-tag positive control lysate, and non-reducing sample buffer for SDS-PAGE. Protocols are provided for both manual and automated magnetic separation workflows. Sufficient components are provided to perform 40 IP or co-IP assays.

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. The Pierce Magnetic HA-Tag IP/Co-IP Kit uses a specific, high-affinity immobilized antibody (clone 2-2.2.14) for rapid purification of HA-tagged fusion proteins from bacterial and mammalian cell lysates, as well as from lysates prepared with the Pierce Human in vitro Translation Kits. The beads are incubated with a cell lysate containing HA-tagged protein, the fusion protein is captured, and the beads are subsequently washed and then eluted using low-pH elution buffer or non-reducing sample buffer. The protocol and buffers have been optimized for both IP and co-IP reactions, enriching for specific protein interaction complexes in the eluted samples. Anti-HA antibody can be used to detect HA-tagged protein by Western blot analysis.

For information about the binding capacity and other properties of the magnetic beads used in this kit, see Pierce Anti-HA Magnetic Beads (Part No. 88836).

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Pierce™ HA-Tag IP/Co-IP Kit