• Product NameRecombinant Bifunctional ligase/repressor BirA, His
• Host SpeciesEscherichia coli
• Purification> 95 % by SDS-PAGE analyses.
• Biological ActivityMeasured by its ability to generate pyrophosphate from the biotinylation reaction. The pyrophosphate is subsequently hydrolyzed using Recombinant Yeast Inorganic Pyrophosphatase/PPA1 (ryPPA1). The specific activity is > 10.0 pmol/min/μg, as measured under the described conditions.
• EndotoxinLess than 0.1 EU/ugof rBirA, His as determined by LAL method.
• Calculated MW Approximately 36.4 kDa, a single non-glycosylated polypeptide chain containing 329 amino acids, with 6 x His at the C-terminus.
• Target Sequence MKDNTVPLKL IALLANGEFH SGEQLGETLG MSRAAINKHI QTLRDWGVDV FTVPGKGYSL PEPIQLLNAK QILGQLDGGS VAVLPVIDST NQYLLDRIGE LKSGDACIAE YQQAGRGRRG RKWFSPFGAN LYLSMFWRLE QGPAAAIGLS LVIGIVMAEV LRKLGADKVR VKWPNDLYLQ DRKLAGILVE LTGKTGDAAQ IVIGAGINMA MRRVEESVVN QGWITLQEAG INLDRNTLAA MLIRELRAAL ELFEQEGLAP YLSRWEKLDN FINRPVKLII GDKEIFGISR GIDKQGALLL EQDGIIKPWM GGEISLRSAE KLEHHHHHH
• Formulation Supplied as a 0.2 μm filtered solution in 50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 10 % glycerol.
• Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 6 months from date of receipt, -20 to -70 ˚C as supplied.- 3 months, -20 to -70 ˚C under sterile conditions after opening.

BirA, the biotin-protein ligase (BPL) of Escherichia coli, is also known as biotin operon repressor, biotin-[acetyl-CoA-carboxylase] ligase, and biotin-[acetyl-CoAcarboxylase] synthetase. BirA, a member of the group II biotin-protein ligase family, contains an N-terminal helix-turn-helix DNA-binding domain, a catalytic core that catalyzes biotinyl 5' adenylate (bio-5'-AMP) synthesis, and a C-terminal domain that plays a role in DNA binding, dimerization, and catalytic function. BirA functions both as a DNA-binding protein that represses the biotin biosynthesis operon as well as an enzyme that synthesizes its own corepressor, bio-5'-AMP, an intermediate in biotinylation reactions. BirA biotinylates via the lysine side chain of biotin-accepting proteins/peptides, including natural substrate, carboxyl carrier protein (BCCP),and Avi Tag fusion proteins. Once biotinylated, (strept)avidin-biotin interactions can be utilized in a wide variety of applications of biochemistry and cell biology, including protein capture, immobilization, multimerizing, and bridging molecules.

• Flow Cytometry (FCM) General Protocol
• Immunofluorescence(IC)
• Immunohistochemistry(Polymer)
• Western Blotting