Kit supplied as:
• Ten (10) vials of 10 µL each 3mg/mL BirA biotin-protein ligase (300 µg total)
• Ten (10) vials of 1.5 mL each BiomixA (0.5 M bicine buffer solution, pH 8.3)
• Ten (10) vials of 1.5 mL each BiomixB (100 mM ATP, 100 mM Mg2+acetate, 500 µM d-biotin)
• Ten (10) vials of 1.5 mL each additional BIO-200 (500 µM d-biotin)
The BirA biotin-protein ligase (EC 6.3.4.15) adds d-biotin covalently
to biotin-acceptor peptides/proteins via an ATP intermediate (biotinyl
5’-adenylate) in a highly efficient and targeted manner. The downstream
applications of enzymatically biotinylated proteins are varied,
important and powerful. The well known biotin-avidin/streptavidin
interaction is often exploited for affinity chromatography or protein
immobilization on surfaces or substrates. Protein detection via
anti-biotin antibodies or avidin/streptavidin-reporter enzyme conjugates
(-HRP, -alkaline phosphatase) or fluorescent probes becomes possible.
Multimeric forms of biotinylated MHC molecules are popular tools in
immunobiology.
When used in combination with our AviTagTM biotin-acceptor
peptide amino acid sequence, biotinylation occurs at twice the rate of
the natural E. coli BCCP substrate and as much as an order of magnitude
or more over other Biotinylation of Peptide sequences. The AviTagTM
sequence consequently requires less of the BirA enzyme and shorter
incubation times to biotinylate to completion than do other sequences
available. If protein instability or protease activities are a concern
this may be important to downstream success.
Our BirA enzyme is E. coli wild-type, encoded by the birA gene, and purified to greater than 99% purity by traditional methods.
Other names for this enzyme include: biotin ligase; biotin operon
repressor protein; birA; biotin holoenzyme synthetase;
biotin-[acetyl-CoA carboxylase] synthetase;
biotin-[acetyl-CoA-carboxylase] ligase; biotin-[acetyl-CoA carboxylase]
synthetase; acetyl CoA holocarboxylase synthetase; acetyl CoA
holocarboxylase synthetase; biotin:apocarboxylase ligase; biotin
holoenzyme synthetase; HCS.
