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形态: 20mM Bicine, 50% Glycerol, pH8.3;
分子量: 35 kDa (Reducing);
纯度: >95% by SDS-PAGE and HPLC;
来源: E.coli;
序列: <p>Met1-Lys321</p>
<p>MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGDACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK.</p>
Component:
Items | Compositon | Amount | |
BirA | 20mM Bicine, 50%Glycerol, pH8.3 | 200ug | 1mg |
10×Buffer A | 500mM Bicine, pH8.3 | 2.5mL | 12.5mL |
10×Buffer B | 100mM ATP, 100mM MgOAc, 500uM D-Biotin | 2.5mL | 12.5mL |
10×Buffer C | 500uM D-Biotin | 2.5mL | 12.5mL |


BirA, the most characterized biotin-acetyl-CoA-carboxylase ligase, has been successfully applied to biotin labeling with a wide range of protein substrates that fused an acceptor peptide (AP, also known as Avitag) in vivo and in vitro. In the presence of ATP, BirA activates biotin to form the BirA-biotinyl-5'-adenylate (BirA-bio-5'-AMP or holoBirA) complex. BirA can either transfer the biotinyl moiety to the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase. Due to its extremely high strength, the interaction between biotin and streptavidin has been exploited for biotechnological applications, like in vivo imaging, cell sorting, et al.

参考图片
Note: 10nmol Avi-tagged protein substrate were labeled by 2μg BirA at 37℃ for 30min. All sample lanes are loaded 4μg protein, respectively. Streptavidin has 4 biotin-binding sites, and may associate with 1 to 4 chains of the biotinylated protein. Therefore, the extent of protein biotinylation can be calculated based on the ratio of biotinylated protein to original substrate protein.
2μg(N: non-reducing condition, R: reducing condition).