Metabolic Biotinylation on Cell Surface

Biotin (vitamin H) is a small molecule present in all living cells, covalently bound by carboxylases and decarboxylases as a coenzyme. Biotin ligases post-translationally covalently link the biotin moiety to specific lysine residues in the BAP sequence in these enzymes. One of the strongest known interactions exists between biotin and avidin, with a dissociation constant of 10^-15 M. This high affinity can resist pH changes and has strong stability. The specificity of biotin and protein binding has a wide range of applications, including protein detection, antigen and nucleic acid analysis, drug targeting, targeted in vivo imaging, and immunotherapy. Facilitated by endogenous biotin ligase, biotinylation can be efficiently performed and biotin can be presented on the cell surface. Alfa Chemistry has an experienced team with many years of experience in the field of high-quality biotinylation, which can provide customers with efficient biotinylation services.

The Services

Currently, researchers have developed a variety of efficient methods and means to express metabolically biotinylated cell surface receptors and enable visualization by efficiently labeling them. Such imaging of cells in vivo will be a useful tool for a wide range of applications in the field of biodiagnostics. In mammalian cells, recombinant proteins can be efficiently biotinylated and display biotin on the cell surface. By combining the labeled streptavidin, it can be detected efficiently. For the biotinylation detection service on the cell surface, the related services that Alfa Chemistry researchers can provide include the following:

• Fluorescence imaging and detection of this habit on the cell surface
• Cell surface staining and detection
• Western blot analysis
• Cell sample culture and processing
• Solution design and development

The visual detection of biotin on the cell surface is of great significance for the detection of biotinylation of recombinant proteins. In the future, more methods will be developed for efficient detection of metabolic biotinylation, which will become an important research tool in molecular biology and cell biology.

Biotin-avidin system for imaging detection

Biotin can be combined with avidin (streptavidin) with high strength. This system can be combined with a variety of imaging reagents (fluorescent dyes, magnetic resonance, PET and SPECT reagents), and has been successfully applied to proteins, nucleic acids and lipids. Quality detection and protein purification. In immunoassays, dyes used to localize or quantify analytes can be coupled via a biotin-avidin system.

Biotin-avidin as an efficient detection system

The biotin-avidin system is suitable for use in a wide range of immunoassays, usually by coupling antibodies to biotin, avidin and other biomolecules such as luciferin and enzymes. The advantage of signal amplification based on biotin-avidin enables the detection signal to be greatly increased and its sensitivity to be significantly improved.

Cell surface biotinylation for protein quantification

The biotin-avidin system is used to chemically label cell surface proteins for direct and accurate cell surface expression, endocytosis and recycling of various plasma membrane receptors and antigens. Quantification of biotin was performed using Western blotting.

In Alfa Chemistry, experts will design a detection system based on the biotin-avidin system for efficient protein detection and analysis according to the characteristics of the sample. If you are interested in our services, please contact us immediately.

Reference:

1. Bakhos. A. T.; et al. Metabolic biotinylation of cell surface receptors for in vivo imaging. Nat Methods. 2006, 3(5): 391-396.

※ It should be noted that our service is only used for research, not for clinical use.