The Avi tag is a short peptide consisting of 15 amino acids (GLNDIFEAQKIEWHE). In the presence of ATP and biotin, BirA attaches a biotin to the lysine residue of the Avi tag, thus achieving biotin labeling of the Avi-tagged target protein.
Beyotime's Biotin Labeling Kit for Avi-tag Protein with BirA (P0630) is designed for fast and efficient biotin labeling of Avi-tagged fusion proteins or peptides by biotin ligase (BirA). BirA efficiently activates biotin (D-Biotin) to form biotinyl-5'adenosine in the presence of ATP and specifically attaches biotin to Avi-tagged lysine residues, enabling fast and efficient biotin labeling of target proteins (Figure 1).
Figure 1. The schematic diagram of Biotin Labeling Kit for Avi-tag Protein with BirA (P0630).
Product Features:
▶ The biotin ligase (BirA) in this kit is derived from E. coli, with high purity and enzyme activity. It does not contain any tags, which does not affect the subsequent detection and purification of biotin-labeled proteins.
▶ The Avi tag is small and almost has no effect on the function of fusion protein. BirA catalyzes the site-specific biotin labeling of Lys residue on the Avi tag, with high efficiency and reproducibility.
▶ The labeling can be performed both in vivo and in vitro, and the labeled protein has a high affinity for streptavidin, thus allowing the Avi-tag technology to be applied to the immobilization and adsorption, purification and detection of the target protein.
▶ Compared with conventional biotin chemical labeling of non-specific sites, BirA-catalyzed reaction conditions are milder and simpler, with less impact on the activity of the labeled protein. Additionally, BirA-catalyzed labeling reaction has high labeling efficiency and specificity.
Typical Figure:
Figure 2. The in vitro labeling of Avi-tagged protein with the Biotin Labeling Kit with BirA (P0630)
The protein (peptide) substrate for this kit is an Avi-tagged fusion protein or peptide. Beyotime has previously released a series of Avi-tagged prokaryotic/eukaryotic expression plasmids that can express specific target proteins in mammalian cells and E. coli. Using these products, biotin labeling of target proteins can be efficiently accomplished in vivo or in vitro.
Eukaryotic Vectors:
| Vector Name | Resistance | Features |
| pCMV-N-3X Flag-Avi-SUMOEU1-Neo | Ampicillin + Neomycin | Containing 3X Flag tag, Avi tag, and SUMOEU1 tag. SUMO tag can be specifically recognized and cleaved by the corresponding protease |
| pCMV-N-3X Flag-SUMOEU1-MCS-Avi | Ampicillin + Neomycin | Containing 3X Flag tag, SUMOEU1 tag, and Avi tag. SUMO tag can be specifically recognized and cleaved by the corresponding protease |
| pCMV-N-3X Flag-Avi-SUMOEU1-MCS-IRES-BirA | Ampicillin + Neomycin | For co-expression of BirA and the target protein containing 3X Flag tag, Avi tag, and SUMOEU1 tag |
| pCMV-N-3X Flag-SUMOEU1-MCS-Avi-IRES-BirA | Ampicillin + Neomycin | For co-expression of BirA and the target protein containing 3X Flag tag, SUMOEU1 tag, and Avi tag |
* The above plasmids contain the ORF of Flag tag and the expressed target proteins can be purified by Anti-Flag Magnetic Beads (P2115/P2181) or Anti-Flag affinity gels (P2271/P2282). Biotin-tagged proteins can be immunoprecipitated by BeyoMag™ Streptavidin Magnetic Beads (P2151), or examined using HRP-labeled Streptavidin (A0303) or AP-labeled Streptavidin (A0312) for in situ detection or immunoblotting.
Prokaryotic Vectors:
| Vector Name | Resistance | Features |
| pET-N-His-Avi | Kanamycin | His tag and Avi tag are located in front of the multicloning site. |
| pET-His-MCS-Avi | Ampicillin | The multicloning site is located between His tag and Avi tag. |
| pET-N-Avi-His-SUMO3 | Ampicillin | Avi tag, His tag and SUMO3 tag are located at the N-terminal of fusion protein. SUMO tag can be specifically recognized and cleaved by the corresponding protease. |
| pET-N-His-SUMO3-Avi | Ampicillin | His tag, SUMO3 tag and Avi tag are located at the N-terminal of fusion protein. SUMO tag can be specifically recognized and cleaved by the corresponding protease. |
| pET-Dual-His-MCS-Avi-BirA | Ampicillin | For co-expression of target protein and BirA under the induction of IPTG. The expressed protein contains His tag at the N-terminal and Avi tag at the C-terminal. The BirA is fused with S tag |
| pET-Dual-Avi-His-SUMO3-MCS-BirA | Ampicillin | For co-expression of target protein and BirA under the induction of IPTG. The expressed protein contains Avi, His, and SUMO3 tags. The BirA is fused with S tag |
| pET-Dual-His-SUMO3-Avi-MCS-BirA | Ampicillin | For co-expression of target protein and BirA under the induction of IPTG. The expressed protein contains His, SUMO3, and Avi tags. The BirA is fused with S tag |
*The above plasmids contain the ORF of His-tag, and the target proteins can be purified using the BeyoGold™ His-tag Purification Resin (P2210/P2218/P2220), His-tag Purification Kit (P2226), BeyoGold™ His-tag Purification Resin (denaturant-resistant) (P2233),His-tag Protein Purification Kit (denaturant-resistant) (P2229S), and BeyoMag™ Anti-His Magnetic Beads (Anti-His Magnetic Beads) (P2135).
Ordering Information