Description
Product Usage Information
Western Blotting: 1:1000
Immunoprecipitation: 1:50
Immunofluorescence (Immunocytochemistry): 1:200 - 1:400
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry)
Specificity / Sensitivity
3X DYKDDDDK Tag (E2T2J) Mouse Monoclonal Antibody (Binds to same epitope as Sigma-Aldrich Anti-FLAG M2 antibody) detects exogenously expressed proteins containing the DYKDDDDK epitope tag. This antibody recognizes the DYKDDDDK peptide, which is the same epitope recognized by Sigma-Aldrich Anti-FLAG M2 antibody, fused to either the amino or carboxy terminus of targeted proteins in transfected cells. This antibody detects exogenously expressed DYKDDDDK-tagged proteins in cells expressed under a CMV promoter. Expression under other promoters has not been evaluated. This antibody detects several non-specific bands of unknown origin in rat cell lines.
Species Reactivity: All Species Expected
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide containing the 3X DYKDDDDK epitope.
Background
Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein's biochemical properties. The DYKDDDDK peptide has been used extensively as a general epitope tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion (1).
