Detail information of Fusion protein Antibodies (His, Myc, HA, GST, MBP, VSV, Thioredoxin, Beta-Gal, GFP)

ADI has produced antibodies to a number of common fusion-tags to study expression of recombinant fusion proteins. In addition, we also provide ready to use affinity matrix (KLH and E.Coli proteins-Sepharose) for the removal of antibodies to KLH or E.coli proteins.

Recombinant Fusion Tags Information

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, 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 (6 x His) 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.

Fusion-tags are finding other ingenious application as well. Peptide can be made with 6X His and then noncovalently bound to Nickel-Sepharose. The Sepharose-bound peptide can be injected to generate antibodies. This method does not require covalent coupling of peptides to carrier proteins. There is a minimum modification of the peptide. In addition, there are no extraneous antibodies to carrier proteins. Conceivably, these tags can also be utilized to monitor interaction of a peptide with other proteins, incubation with live cells or even develop competitive immunoassays.