ANTI-FLAGantibody, Rat monoclonal

Uses

ANTI-FLAG^? antibody, Rat monoclonal has been used in:

• chromatin immunoprecipitation (ChIP)
• western blotting
• coimmunoprecipitation
• flow cytometric analysis

General Description

Monoclonal Anti-FLAG^? (rat IgG1 isotype) is derived from the hybridoma 6F7 produced by the fusion of mouse myeloma cells and splenocytes from rat immunized with the FLAG^? peptide. The antibody is purified from culture supernatant of hybridoma cells grown in a bioreactor.

Monoclonal Anti-FLAG^? recognizes N-terminal,
C-terminal and internal Flag-tagged fusion proteins. The product is especially recommended for identifying C-terminal FLAG^?-tagged fusion proteins.
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 associationith 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 of the C-terminal FLAG^? peptide from a fusion protein catalyzed by Cu²⁺ ions 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 b-phosphatase, as well as in the human and bovine enzyme.