DescriptionE coli BirA acts both as a biotin-operon repressor and as the enzyme that synthesizes the corepressor, acetyl-CoA:carbon-dioxide ligase. This protein also activates biotin to form biotinyl-5'-adenylate and transfers the biotin group to the amino group of a lysine residue present within biotin-accepting proteins, such as biotin carboxy carrier protein (BCCP) or proteins fused to a 15 amino acid peptide termed AviTag (GLNDIFEAQKIEWHE)
Applications

Covalently biotin-labeling of acceptor proteins fused to AviTag in vitro for detection and purification assays

SourcePurified from a recombinant E. coli strain carrying the E. coli birA protein ligase gene (NP_418404)
ComponentsBirA Biotin Ligase, 1 μg/μl in storage buffer: 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT, 50% glycerol, pH 7.5 @ 25°C.
Activity1 μg is sufficient to transfer biotin into 50μg target protein at room temperature or at 4°C overnight (>95%).
Quality Control
  • The absence of protease was confirmed by appropriate quality tests.
  • Functionally tested for the capacity to label proteins with biotin in vitro.
Storage-20°C.