POLYCLONAL ANTI-FLAG-PEROXIDASE ANTIBODY PRODUCED IN RABBIT

General description

Anti-FLAG antibody is developed in rabbits using purified FLAG fusion protein as immunogen. Whole antiserum is purified using protein A immobilized on agarose to provide the IgG fraction of the antiserum and is conjugated to horseradish peroxidase.

ANTI-FLAG recognizes the FLAG epitope located on FLAG fusion proteins. The antibody reacts with N-terminal, N-terminal-Met, and C-terminal FLAG fusion proteins by immunoblotting. Specific staining is inhibited by the FLAG peptide (N-Asp-Tyr-Lys-Asp-AspAsp-Asp-Lys-C). Applications for the conjugate include Western blots and ELISA.

Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance, or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with the FLAG peptide sequence may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, or at internal positions of the target protein. FLAG may also be placed in association with other tags. The small size of the FLAG tag or sequence and its high hydrophilicity tend to decrease the possibility of interference with the protein expression, proteolytic maturation, antigenicity, and function. The N-terminal FLAG peptide sequence contains a unique enterokinase cleavage site allowing it to be completely removed from the purified fusion proteins. Cleavage catalyzed by Cu²⁺ ions of the C-terminal FLAG peptide from a fusion protein has been reported. A sequence motif with five out of eight amino acid residues identical to the FLAG peptide is found in both rat and mouse Mg²⁺ dependent protein β-phosphatase, as well as in the human and bovine enzyme.

Immunogen

purified FLAG fusion protein

Application

For immunoblotting, with 1:2000 – 1:4000 as an initial suggested dilution
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Physical form

Lyophilized powder

Legal Information

FLAG is a registered trademark of Sigma-Aldrich Co. LLC

Disclaimer

For Research use only. Not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.