Nonimmuno Biotinylation to Detect Protein Interactions

Proteins control all biological systems in cells. In addition to completing their own functional tasks independently, most proteins need to interact with other proteins to ensure complete biological activity. Detecting protein interactions in cells is critical to understanding protein biology. The most commonly used detection methods for protein interactions are co-immunoprecipitation (co-IP), pull-down detection, cross-linking, label transfer, and Far-Western blot analysis. Currently employed detection methods require the use of specific antibodies for immunoprecipitation and detection, making them costly and unreliable. At present, researchers have successfully avoided the impact of the specificity of specific antibodies, and can detect the interaction between proteins through a non-immune method, which has the advantages of low price, sensitivity and reliability. Alfa Chemistry's researchers have long been committed to high-quality biotinylation services, and meet customer needs with a wide range of strategies. We can meet the needs of customers in terms of time, specifications and prices, and can design suitable biotinylation reactions for specific samples to continuously meet customer needs.

Proteins play an important role in the functions of organisms, and the ability to efficiently detect protein interactions plays an important role in mastering biology. Alfa Chemistry has a professional biotechnology platform, which can provide reasonable biotinylation solutions and strategies, professional methods and means, and efficient biotinylation services to meet customer needs. Related services we can provide include but are not limited to:

• Biotinylated protein interaction assays
• Cell co-sedimentation analysis
• Calculation of binding affinity

Current research

To achieve efficient testing of protein interactions is of great significance to the study of proteomics. Currently, researchers have developed a new detection platform Avitag-BirA for efficient detection of protein-protein interactions. After cells are lysed, the protein encoding Avitag is removed by Glutathione Sepharose preloaded with GST fusion protein. The interaction of biotinylated Avitag protein and GST fusion protein enables efficient detection.

Figure 2. Scheme depicting the principles of protein-protein interaction assays using the Avitag-BirA system

About protein interactions

Protein-protein interactions are linked to numerous biological processes, including signal transduction, tissue integrity and force generation. Protein interaction is usually achieved by co-immunoprecipitation, which is costly and unreliable. The biotin-avidin/streptavidin system is one of the most potent currently known. BirA is an E. coli biotin ligase that can be used for protein purification, detection and fluorescence imaging through specific conjugation. BirA-mediated biotinylation system detects protein-protein interactions without the use of specific antibodies.

The wide application

• Determine interactions with membrane proteins and other peptides
• Screening of protein-protein interactions
• Construction of protein network system
• Screening for interactions or transient interactions

Alfa Chemistry focuses on providing efficient non-immune biotinylation services, and can provide customized biotinylation services according to customer sample requirements. If you are interested in our services, please contact us immediately.

Reference:

1. Huang. C.; et al. Detection of protein-protein interactions using nonimmune IgG and BirA-mediated biotinylation. Biotechniques. 2010, 49(6): 881-886.

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