A fusion tag called FLAG consists of eight amino acids (AspTyrLysAspAspAspAspLys) including an enterokinase-cleavage site. FLAG is specifically designed for immunoaffinity chromatography. It allows elution under non-denaturing conditions. Several antibodies against this peptide have been developed. One antibody denoted as M1 binds the peptide in the presence of bivalent metal cations preferably Ca(+). Elution is effected by chelating agents. Another strategy is competitive elution with excess of free Flag Peptide. The FLAG peptide purifies and detects recombinant fusion proteins. FLAG peptide is useful in Western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization. Flag peptide is a hydrophilic tag which significantly improves the detection and purification of recombinant fusion proteins.
Anti-human FLAG mAb, is derived from hybridization of mouse F0 myeloma cells with spleen cells from BALB/c mice immunized with a Synthetic peptide (DYKDDDDKC)-KLH.
Mouse IgG1 heavy chain and k light chain.
FLAG antibody was purified from mouse ascitic fluids by protein-A affinity chromatography.
1mg/ml containing PBS, pH-7.4, 10% Glycerol and 0.02% Sodium Azide.