BirA Biotin Protein Ligase, Recombinant
BirA Biotin Protein Ligase, Recombinant
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Recombinant BirA Biotin Ligase (EC 6.3.4.15)
BirA (also known as biotin protein ligase or BPL) is widely used for antibody biotinylation, proximity-dependent biotinylation (BioID), and streptavidin/avidin-based pull-down assays. Because biotinylation is confined to the AviTag sequence (GLNDIFEAQKIEWHE), it avoids the non-specific labeling seen with chemical NHS-ester reagents, preserving protein activity and enabling oriented capture on streptavidin surfaces. Unlike chemical NHS-ester methods that produce heterogeneous mixtures, BirA achieves site-specific biotinylation exclusively at the AviTag lysine residue.
Key Features & Specifications:
- Source & Expression: Expressed in E. coli cell expression system (amino acids 3–321).
- Molecular Weight: 36.2 kDa.
- Purification Tag: Features an N-terminal His-tag, allowing for easy removal post-reaction using a Ni-NTA column (if the substrate lacks a His-tag).
- Purity: >90% as determined by SDS-PAGE (Coomassie staining).
- High Activity: 5,000 Units/µg. (1 Unit biotinylates 1 pmol of AviTag peptide substrate in 30 minutes at 30°C).
- Concentration: 1.0 mg/mL (by A280).
Applications:
- Site-specific in vitro biotinylation of target proteins and antibodies.
- Proximity-dependent biotinylation and cell labeling applications.
- Preparation of high-affinity complexes for streptavidin/avidin pull-downs, immobilization, and functionalization assays.
Catalog # BBL-301
This product is intended for laboratory research use only.
Expression system: E.Coli
Amino acids: 3-321
Tag(s): N-terminal His-tag
Molecular weight: 36.2 kDa
Storage buffer: 50 mM Tris-HCl, pH 7.5, 50 mM NaCl, 3 mM DTT, and 50% Glycerol
Concentration: 1.0 mg/mL by A280
Purity: >90% (SDS-PAGE, Coomassie staining)
Activity: 5,000 Units/µg
Definition of Activity: 1 Unit is the amount of enzyme that will biotinylate 1 pmol of AviTag peptide substrate in 30 minutes at 30°C in the reaction buffer containing 10 mM Tris-HCl, pH 8.0, 10 mM ATP, 10 mM MgOAc, 50 µM d-Biotin, and 38 µM peptide substrate.
Storage is recommended at -20°C for longer periods of time
Product Description and Manual
Shipping: Product requires shipping on dry ice. Please contact info@amidbiosciences.com for shipment estimates.
References:
- Mark Howarth & Alice Y Ting. Imaging proteins in live mammalian cells with biotin ligase and monovalent streptavidin. (2008) Nature Protocols 3: 534
- Askin et al. Green fluorescent protein-based assays for high-throughput functional characterization and ligand-binding studies of biotin protein ligase. (2016) Anal. Methods 8: 418
- Cai Huang and Ken Jacobson. Detection of protein-protein interactions using nonimmune IgG and BirA-mediated biotinylation. (2010) BioTechniques 49: 881
- de Boer et al. Efficient biotinylation and single-step purification of tagged transcription factors in mammalian cells and transgenic mice. (2003) Proc. Natl. Acad. Sci. USA 100: 7480
- Elif Nur Firat-Karalar, Tim Stearns. Probing mammalian centrosome structure using BioID proximity-dependent biotinylation. (2015) Methods in Cell Biology, 129: 153
Domestic Shipping in the US
Temperature sensitive products requiring shipping/handling on dry ice are shipped via UPS Next Day Air Saver. These products are delivered the next day if your order is placed before 3 PM PST. A dry ice surcharge is included in the shipping cost at checkout. For orders totaling over $1,000, free shipping and handling will be applied to the order.
International Shipping
Orders to be shipped internationally (outside of the US) can be shipped via DHL Express Worldwide or FedEx International Priority. Please contact info@amidbiosciences.com if you would like your company's shipping account to be used for the order. Our customers have sometimes negotiated lower shipping rates than us.
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