BirA-transformed Competent E. coli cells
BirA-transformed Competent E. coli cells are supplied as 10 x 50 µl vials in aqueous solution containing 10 mM Na MOPS, pH 7.0, 75 mM CaCl2, 10 mM KCl and 15% glycerol. Strain BL21(DE3), a chemically competent E. coli B strain, contains an IPTG-inducible BirA expression plasmid and a constitutively expressed streptomycin/spectinomycin resistance gene. These cells are compatible with most cloning vectors and are suitable for expression and biotinylation of recombinant proteins using AviTag™ technology.
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Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
- E coli in vivo Biotinylation Kit (without BirA Competent Cells) (BPS Bioscience #78870)
- SOC or LB medium
- Expression vector containing gene of interest and the AviTag™ sequence
- Streptomycin or spectinomycin, and appropriate antibiotic based on expression vector for the gene of interest
- Water bath
- Protease inhibitor cocktail (for example, protease cocktail from Sigma1 #P8465: AEBSF 23 mM, EDTA 100 mM, Bestatin 2 mM, Pepstatin A 0.3 mM, E-64 0.3 mM).
- Affinity chromatography for AviTag™
| Catalog # | Name | Amount | Storage |
| 27462 | Bir-A transformed Competent E. coli cells | 10x 50 µl | -80°C |
Biotin-labeling is commonly used for non-radioactive labeling and purification of proteins or other target molecules. Biotin labeling takes advantage of the exceptionally strong interaction between biotin (vitamin H) with avidin or streptavidin. The affinity of biotin to avidin or streptavidin is one of the strongest known non-covalent interactions known between a protein and a ligand, exhibiting a dissociation constant (Kd) around 4x10-14 M. BirA, an E. coli biotin protein ligase, covalently adds biotin to an AviTag™ peptide in a highly efficient and specific manner, in a reaction that requires ATP. It has the advantage of resulting in a homogeneous product that can be used in protein capture, immobilization and functionalization.
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