This antibody was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the termini to facilitate coupling.
The purified antibody is directed against DYKDDDDK and is useful in determining its presence in various assays where the epitope tag is present at either the amino or carboxy terminus of recombinant proteins. This monoclonal antibody detects over-expressed proteins containing DYKDDDDK. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.
Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein's biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells.
Sterile Solution 0.02 M Potassium Phosphate, pH 7.2, 0.15 M NaCl, 0.01% Sodium Azide
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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