Anti-Hemagglutinin Tag (HA Tag) (Agarose)

Expression of genes in E. coli or yeast or baculovirus offers a convenient system to produce large amounts of recombinant proteins that may otherwise be difficult to isolate from natural cells and tissues. Very often antibodies to these newly identified proteins are not available to study its biochemical properties, monitor protein expression, and purification. In order to circumvent this problem, short pieces of well-defined peptides (Poly-His, Flag- epitope or c-myc epitope or HA-tag) or small proteins (bacterial GST, MBP, Thioredoxin, b-Galactosidase, VSV-Glycoprotein etc) are often cloned along with the target gene. Proteins are expressed as fusion proteins. Antibodies to these fusion-tags are already available to monitor fusion protein expression and purification. Therefore, fusion-tags serve as universal tags much like secondary antibodies. Many tags have their own characteristics. Poly-His-fusion proteins (6xHis) can bind to Nickel-Sepharose or Nickel-HRP. GST-fusion proteins can bind to glutathione-Sepharose. Therefore, a high degree of purification of fusion protein can be achieved in just one affinity purification step. Purity of fusion proteins can be followed by Tag-antibodies. Very often, fusion proteins are directly injected into animals to generate antibodies. Some fusion tags can be removed later by treatment with enzymes to generate tag-free recombinant proteins. Applications: Suitable for use in Immunoprecipitation and for affinity purification of fusion proteins containing tag. Other applications not tested. Recommended Dilution: Immunoprecipitation: 25-50ul of gel volume (50-10ul of 1:1 suspension) per 500ug of the protein lysate., Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.