Thermo Scientific Pierce Anti-HA Agarose is a high-affinity immunoprecipitation resin ideal for purification of recombinant HA-tagged proteins expressed in human in vitro expression systems and bacterial and mammalian cell lysates.
The anti-HA antibody used to manufacture Pierce Anti-HA Agarose is a highly specific mouse IgG1 monoclonal antibody that recognizes the HA-epitope tag (YPYDVPDYA) derived from the human influenza hemagglutinin (HA) protein. This anti-HA affinity resin can be packed into gravity purification columns, spin purification columns or cartridges for FPLC instruments to purify HA-fusion proteins expressed in bacterial or mammalian cells.
Highlights:
- Specific – highly specific anti-HA monoclonal antibody enable high-yield and high-purity for immunoprecipitation
- Scalable – available in 1 and 5mL resin package sizes to allow for larger scale purifications or immunoprecipitations
- Versatile – can be used in spin or gravity columns as well as in FPLC cartridges
- Convenient – reagents to elute and detect HA-tagged fusion proteins are available separately
Applications:
- Purification of HA-fusion proteins expressed in the Pierce Human In Vitro Translation Kits
- Large scale purification of recombinant HA-tagged proteins
- High-throughput enrichment of fusion proteins and interacting partners
Product Details:
Pierce Anti-HA Agarose has a highly specific monoclonal anti-HA antibody that is covalently immobilized on a crosslinked 4% beaded agarose support. The product is supplied as 50% slurry. Upon incubation with a sample, HA-tagged fusion protein is captured on the agarose beads. After simple washing steps, the tagged protein easily eluted from the resin using 0.1M Glycine, pH 2.0-2.8, 3M NaSCN or 50mM NaOH depending on the downstream application of the purified protein. For elution of highly functional proteins, Pierce HA-peptide can also be used to elute the HA-tagged protein. Anti-HA antibody may be used to detect the presence of the tagged protein by Western blot detection.
 |
Immunoprecipitation of HA-tagged protein (GST-PI3K-SH2-HA) from E. coli lysate 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. |
 |
Purification of HA-tagged protein from the Pierce human in vitro expression lysate. HA-tagged GFP (from Pontellina plumata) was expressed with the Pierce Human In Vitro Protein Expression System and purified by incubation with Pierce Immobilized Anti-HA Agarose. Anti-HA agarose slurry (50µL) was added to a Pierce Spin column (Part No. 69705) along with 60µL of the in vitro translation reaction diluted to a final volume of 200µL in TBS. The resin and sample were mixed for 1 hour at 4°C with end-over-end mixing. The resin was pelleted by centrifugation and was washed 3X with 10 column volumes of TBS-T. HA-tagged GFP was eluted from the resin by adding 3 x 1 column volume of 1mg/mL Pierce Influenza Hemagglutinin (HA) Peptide (Part No. 26184). The elution fractions were combined and 20% of the elution fraction was electrophoresed in SDS-PAGE and stained with Gel Code Blue (Part No. 24590). Lane 1: immunoprecipitation eluate from human in vitro translation reaction containing no DNA (negative control), Lane 2: immunoprecipitation eluate from human in vitro translation reaction containing GFP-HA DNA, Lane 3: MW Marker |
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:
IgG Elution Buffer
Anti-HA Antibody
Other Anti-Tag Antibodies
HA-Tag IP/Co-IP Kit
Fusion Protein Purification Resins and Kits
Cell Lysis and Protein Extraction Reagents
|
