Creative BioMart has developed its own protein biotinylation platform for more than 10 years, and has completed a variety of customized protein biotinylation services for customers in the field of academic research and industrial production.
Figure 1. Protein Biotinylation (Man, 2021)
Biotin labeling is very useful in protein research and can be used in a variety of applications, including enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. It can also be used in immunohistochemistry, immunoprecipitation, cell surface labeling and flow cytometry. In addition, there is a very strong indication that biotinylation can also be used to examine protein-protein interactions and characterization in living cells.
Biotin labeling in vitro is simple and efficient, and can be completed without changing the biological activity of the target protein. Biotinylated proteins are immobilized on streptavidin-coated surfaces/beads and biotinylated by enzymatic or chemical methods.
If necessary, we can monitor the biotitization of proteins in three different ways (1) enzyme-linked immunosorbent assay (ELISA) is a method to prove that the target protein has been biotized rapidly; (2) Use a kit to determine the biotin quantity per protein molecule; (3) The presence of abiotic protein is evaluated by the combination of SDS-PAGE in the samples with the magnetic beads coated by streptomycin.
The above protein biotinylation services are applicable to recombinant proteins produced in bacteria, insect cells, mammalian cells, plants and other expression systems.
In addition to the in vitro biotinylation service, we have developed an effective in vivo biotin program. This scheme is mainly suitable for the E. coli expression system. BtAP is the substrate of E.coli biotin holoenzyme synthetase (BirA), which is a biotinylated enzyme of E.coli biotin carboxyl carrier protein (BCCP). We have developed an efficient bacterial expression vector, which can express the target protein and BirA gene fused with BtAP under the bacterial operon. Using this vector and the optimized bacterial host strain, the target protein of BTAP fusion is co-expressed with BirA, and the target protein will be completely biotinylated in vivo.
Figure 2. Biotinylation experiments (Twelvetrees, 2013)
Biotinylation is an ideal choice for protein purification for many reasons. Here are some of them.
Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.
References
1. Man, T., 2021. The Advantages of Biotinylation Tagging in Protein Purification. [online]
2. Twelvetrees, Alison (2013): Cell surface biotinylation experiments. figshare.
