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.
With sufficient materials to perform 25 IP or Co-IP assays, the complete HA Tag IP/Co-IP Kit includes the immobilized anti-HA antibody on agarose resin, binding, wash and elution buffers, microcentrifuge spin columns, HA-tag positive control lysate, sample loading buffer and easy-to-follow instructions.
Highlights:
- 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
Applications:
- purification and detection of HA-tagged fusion-protein for downstream assays
- enrichment and screening of HA-tagged fusion-protein mutants
- co-immunoprecipitation and screening of interacting proteins
Product Details:
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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Immunoprecipitation of HA-tagged protein (GST-PI3K-SH2-HA) with the Thermo Scientific Pierce HA-Tag IP/Co-IP Kit. Coomassie Stained gel. Lane 1: MW marker, Lane 2: 15µL of 0.25mg/mL of positive control lysate, Lane 3: 15µL of Flow-through, Lane 4: 15µL of Glycine, pH 2.8 elution fraction |
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| Co-immunoprecipitation of active E2F1:HA-tagged pRb complex. HA-tagged pRb was expressed using a Pierce Human in vitro Expression System. 50µL of the translation reaction was diluted to 200µL in TBS and added to 15µL of agarose resin and incubated for 1 hour at 4°C as a non-specific pre-clearing step. The cleared translation reaction was added to 50µL of anti-HA resin and incubated for 3 hours at 4°C with end-over-end rocking. The resin was washed with 3 x 500µL of TBS-T. HA-pRb:E2F1 complex was eluted with the addition of 30µL of 2X non-reducing sample buffer. In vitro translation reactions without and with HA-pRb DNA (lanes 1 and 3, respectively) and anti-HA agarose elution fractions from each (lanes 2 and 4, respectively) were probed with rabbit anti-E2F1 (Part No. PA5-17549) and mouse anti-HA (Part No. 26183) and detected with Thermo Scientific SuperSignal West Dura Extended Duration Substrate (Part No. 34075). |
References:
- Kolodziej, P.A. and Young, R.A. (1991). Methods Enzymol. 194, 508-519.
- Chen, Y., et al. (1993). Proc. Natl. Acad. Sci. U.S.A. 90, 6508-6512.
- Qoronfleh, M.W., et al. (2003). J. Biomed. Biotechnol. 2003(5), 291-298.
- Brymora, A., et al. (2001). Anal. Biochem. 295, 119-122.
Related Products:
Tag-Based IP and Pull-Down Kits
Immunoprecipitation (IP) Kits
Cell Lysis and Protein Extraction Reagents
Immobilized Anti-HA Resin
Anti-HA Antibody
Other Anti-Tag Antibodies
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