In Vivo Biotinylation to Detect Proteins

There are a large number of protein-protein interactions in the cell, often acting on the whole process of the cell. In gene regulation, proteins can be identified to target DNA sites on chromosomes. In biochemistry, biotin is a useful label on proteins. The high-affinity interaction of biotin with avidin or streptavidin has been widely used in biochemistry and molecular biology. In vivo labeling of proteins by covalent attachment of biotin moieties has emerged as a new promising tool for protein detection and purification. Proteomic approaches require simple and efficient protein detection methods, and researchers have developed highly efficient methods for the detection of protein biotinylation in vivo. Alfa Chemistry provides advanced biotinylation services, focusing on a wide range of biotinylation solutions and strategies to meet customer needs with precise detection methods.

The Services

Alfa Chemistry has a professional biotechnology team equipped with advanced equipment dedicated to high-quality biotinylation platforms. We are committed to optimizing the service system and insisting on providing customers with satisfactory services, including but not limited to:

• In vivo biotinylation detection protein detection
• Sample cultivation, transfection and selection
• Immunofluorescence microscopy test
• Purification of biotinylated molecules
• Protein analysis
• Quantitative analysis
• Determination of biotinylation efficiency
• Site-specific biotinylation

The methods to identify and analyze biotinylated proteins in vivo

• Protein affinity chromatography: Using peptides as baits in affinity pull-down experiments can be used to efficiently detect proteins. Synthetic peptides for competitive disruption of binding can be used to validate protein interactions. Peptides can be tagged with 6His, FLAG, V5 HA or cMYC for protein affinity chromatography.
• Affinity Blotting: Proteins can be fractionated by SDS-PAGE and identified by their ability to bind peptides, proteins or ligands. During blotting, the protein can be recovered by removing the denaturant, and then the protein of interest can be labeled for qualitative analysis.
• Immunoprecipitation: The commonly used method for detecting proteins is co-immunoprecipitation (Co-IP). The analyte is lysed, antibodies are added, immunoprecipitated, washed, and finally analyzed.
• Cross-linking: Using membrane-permeable cross-linking agents in vivo, the cross-linked protein can be released from the complex by cleaving the cross-link. After cross-linking, immunoprecipitation of the ligand protein was performed.

Biotin-streptavidin/avidin system

The efficient characterization of proteins and protein interactions is mainly through the biotin-avidin system, which is widely used in the medical field. The biotin-avidin system can be combined with a variety of markers, and the high affinity can be amplified, making the biotin-avidin system more sensitive. The biotin-avidin system has high sensitivity, high specificity, high stability and universality.

Figure 2. The biotin-avidin system can be applied to biological diagnosis, affinity chromatography, localization observation and gene probe

Alfa Chemistry offers comprehensive biotinylation services to meet customers' needs. If you are interested in our services, please contact us immediately.

References:

1. Andrea. P.; et al. In vivo site-specific biotinylation of proteins within the secretory pathway using a single vector system. BMC Biotechnol. 2008, 8: 41.
2. Antoine. V.; et al. Use of protein biotinylation in vivo for immunoelectron microscopic localization of a specific protein isoform. J Histochem Cytochem. 2008, 56(10): 911-919.

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

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