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Recombinant E. coli BirA His-tag Protein, CF

Catalog # 10464-BA | R&D Systems, Inc. a Bio-Techne Brand
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10464-BA-100

Key Product Details

Source

E. coli

Accession #

Conjugate

Unconjugated

Applications

Enzyme Activity

Product Specifications

Source

E. coli-derived e. coli BirA protein
Lys2-Lys321, with an N-terminal Met & 6-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

36 kDa

SDS-PAGE

30-38 kDa, under reducing conditions

Activity

Measured by its ability to generate pyrophosphate from the biotinylation reaction.  The pyrophosphate is subsequently hydrolyzed using Recombinant Yeast Inorganic Pyrophosphatase/PPA1 (ryPPA1) (Catalog # 8088-PP).

The specific activity is >10.0 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant E. coli BirA His-tag Protein, CF

Recombinant E. coli BirA His-tag Protein SDS-PAGE

Recombinant E. coli BirA His-tag Protein SDS-PAGE

2 μg/lane of RecombinantE. coliBirA His-tag (Catalog # 10464-BA) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 30-38 kDa.

Formulation, Preparation and Storage

10464-BA
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl, TCEP, EDTA and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: BirA

BirA, the biotin-protein ligase (BPL) of Escherichia coli, is also known as biotin operon repressor, biotin-[acetyl-CoA-carboxylase] ligase, and biotin-[acetyl-CoA-carboxylase] 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 (1). 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 (1). BirA biotinylates via the lysine side chain of biotin-accepting proteins/peptides, including natural substrate, carboxyl carrier protein (BCCP), and AviTag fusion proteins (1-4). 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 (5).

References

  1. Chapman-Smith, A. and et al. (2001) Protein Sci. 10:2608.
  2. Cronan, J. (1989) Cell. 58:427.
  3. Cull, M. and Schatz, P. (2000) Methods Enzymol. 326:430.
  4. Li, Y. and Sousa R. (2012) Protein Expr. Purif. 82:162.
  5. Fairhead, M. and Howarth, M. (2015) Methods Mol. Biol. 1266:171.

Long Name

DNA-binding transcriptional repressor/biotin-[acetyl-CoA-carboxylase] ligase BirA

Alternate Names

bioR, dhbB, ECK3965

Entrez Gene IDs

948469 (E. coli)

Gene Symbol

BIRA

UniProt

Product Documents for Recombinant E. coli BirA His-tag Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant E. coli BirA His-tag Protein, CF

For research use only

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