This antibody was purified from whole rabbit serum prepared by repeated immunizations with the Enterokinase Cleavage Site (ECS) peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the C-terminus end to facilitate coupling. This antibody reacts with FLAG conjugated proteins.
This affinity purified antibody is directed against DYKDDDDK and is useful in determining its presence in various assays. This polyclonal antibody detects over-expressed proteins containing DYKDDDDK at either the N or C terminus. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins.
This antibody is optimally suited for monitoring the expression of DYKDDDDK tagged fusion proteins. As such, this antibody can be used to identify fusion proteins containing the DYKDDDDK epitope. The antibody recognizes the epitope tag fused to the amino- and carboxy- termini of targeted proteins. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and DYKDDDDK containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation and immunocytochemistry, and other immunodetection techniques. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. Now the most commonly used hydrophilic octapeptide is DYKDDDDK. This polyclonal antibody to detect DYKDDDDK conjugated proteins binds DYKDDDDK containing fusion proteins with greater affinity than the widely used monoclonal M1, M2 and M5 clones, and shows greater sensitivity in most assays. Affinity purification of the polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins. The applications listed have been tested for the unconjugated form of this product. Other forms have not been tested.
0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2, 1% BSA, 0.01% Gentamicin sulfate
Reconstitute in deionized water
Short term: store at 4°C. Long term: aliquot and store at -20°C. Avoid freeze-thaw cycles.
For research use only.
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