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Contributor Information

  • Institute A*STAR Accelerate Technologies Pte Ltd

Tool Details

  • Tool name: Anti-BirA [6C4c7]
  • Alternate names: BirA antibody, bioR antibody, dhbB antibody, JW3941 antibody
  • Clone: 6C4c7
  • Tool type: Antibodies
  • Class: Monoclonal
  • Conjugate: Unconjugated
  • Reactivity: E.coli
  • Host: Mouse
  • Molecular weight of the target: 35 kDa
  • Application: IF ; WB
  • Strain: C57BL/6
  • Description: Monoclonal antibody to investigate the E.coli BirA protein. Background and Research Application E.coli BirA is a well-characterized multifunctional protein that either catalyses the transfer of biotin to biotin carboxyl carrier protein (BCCP), an acetyl-CoA carboxylase subunit or, alternatively, binds to the biotin operator resulting in transcriptional repression of the biotin operon. This protein also activates biotin to form biotinyl-5'-adenylate and transfers the biotin moiety to biotin-accepting proteins. A mutant BirA (R118G) shows loss of DNA binding ability and promiscuity of substrate biotinylation. These properties of BirA R118G were used to develop an in vivo proximity labelling technique that identifies potential interacting proteins (BioID). In the BioID method, a protein of interest is tagged with BirA R118G and expressed in live cells. Addition of the biotin to the culture medium results in biotinylation of proteins in vicinity of the bait. The biotinylated proteins can then be affinity purified with biotin binding proteins such as streptavidin or avidin even under harsh denaturing conditions and subsequently identified by mass spectrometry or immunoblot analysis.
  • Immunogen: GST fused to E.coli BirA R118G
  • Immunogen UniProt ID: P06709
  • Isotype: IgG1 kappa
  • Research area: Tags and cell markers
  • Myeloma used: Sp2/0-Ag14

  • For Research Use Only

Target Details

  • Target: BirA R118G
  • Target molecular weight: 35 kDa
  • Target background: Monoclonal antibody to investigate the E.coli BirA protein. Background and Research Application E.coli BirA is a well-characterized multifunctional protein that either catalyses the transfer of biotin to biotin carboxyl carrier protein (BCCP), an acetyl-CoA carboxylase subunit or, alternatively, binds to the biotin operator resulting in transcriptional repression of the biotin operon. This protein also activates biotin to form biotinyl-5'-adenylate and transfers the biotin moiety to biotin-accepting proteins. A mutant BirA (R118G) shows loss of DNA binding ability and promiscuity of substrate biotinylation. These properties of BirA R118G were used to develop an in vivo proximity labelling technique that identifies potential interacting proteins (BioID). In the BioID method, a protein of interest is tagged with BirA R118G and expressed in live cells. Addition of the biotin to the culture medium results in biotinylation of proteins in vicinity of the bait. The biotinylated proteins can then be affinity purified with biotin binding proteins such as streptavidin or avidin even under harsh denaturing conditions and subsequently identified by mass spectrometry or immunoblot analysis.

Application Details

  • Application: IF ; WB

Handling

  • Format: Liquid
  • Concentration: 1mg/ml
  • Storage buffer: PBS with 0.02% azide
  • Storage conditions: -15°C to -25°C
  • Shipping conditions: Shipping at 4°C

Documentation

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References

  • •  Roux et al. 2012. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells J Cell Biol. 196(6):801-10. PMID: 22412018.
  • •  Choi-Rhee et al. 2004. Protein Sci. 13(11):3043-50. PMID: 15459338.
  • •  Goodchild et al. 2004. Mislocalization to the nuclear envelope: an effect of the dystonia-causing torsinA mutation. Proc Natl Acad Sci U S A. 101(3):847-52. PMID: 14711988.
  • •  Streaker et al. 2003. Coupling of protein assembly and DNA binding: biotin repressor dimerization precedes biotin operator binding. J Mol Biol. 325(5):937-48. PMID: 12527300.
  • •  Kwon et al. 2000. Multiple disordered loops function in corepressor-induced dimerization of the biotin repressor. J Mol Biol. 304(5):821-33. PMID: 11124029.