WB-Based Exosome Verification

Western blotting (WB) is a widely used analytical technique for detecting specific proteins in samples. Since exosomes have a unique protein expression profile, WB has been used to identify the presence and expression level of specific single proteins on the surface of exosomes.

Exosome marker proteins

Exosomes are nano-scale ( typically ~50-100 nm in diameter), cell-derived extracellular vesicles (EVs) released through exocytosis of multivesicular bodies (MVBs) via the endosomal pathway. Thus, exosomes naturally contain components involved in membrane transport and fusion-related processes (such as flotillin, Rab and GTPases) as well as MVB production (Alix and TSG101). Exosomes also encompass heat shock proteins (HSP60, HSP70, HSPA5, CCT2 and HSP90), integrins and tetraspanins (such as CD63, CD81 and CD9), lipid-related proteins, and phospholipases. In addition, other proteins contained in exosomes are associated with the cytoplasmic content of their cellular origin. Consequently, some of these components are frequently utilized as exosome markers in WB analysis. Exosome proteins, such as CD63, CD9, CD81 and TSG101, HSP70, Alix, etc. are the most commonly used exosome vesicle markers.

WB-based exosome verification

Although WB is not the most advanced technique, it is essential for exosome identification and characterization in the current research. With our well-established technologies and experienced experts, Creative Proteomics offers end-to-end WB-based exosome verification services, including exosome protein preparation, antibody selection, WB analysis, and comprehensive analytical report delivery.

According to the guidelines suggested by the International Society for Extracellular Vesicles (ISEV), the investigation of a minimum of one protein from each of the four categories to claim the presence of EVs. Detection of CD63 and flotillin-1 would increase the reliability of the results, as CD63 in particular is often used as a marker for exosomes in the literature. Our team of experts will provide customized solutions through customers' specific projects. We specialize in the improved and robust workflow to support exosome protein identification. By choosing our WB-based exosome verification service, we guarantee that your project meets the highest standards of quality, technical analysis, and reporting.

We can help you

• Exosome size, and size distribution
• Exosome purity, integrity, and internal morphology
• Exosome stability assessment
• Clusters/aggregates and cell debris/impurities in exosome preparation

Features of Our Services

• Improved and robust WB protocol and workflow
• Easy-to-use and good-quality antibodies
• Highly experienced and professional team
• Whole-process technical trace service

Exosomes are nano-scale, cell-derived EVs that are thought to be important mediators of intercellular communication. To better understand the exosomes, it is necessary to confirm and assess each exosome preparation each time to maintain consistent and reproducible quality. With the increasing use of exosomes in diagnostic and pharmaceutical applications, accurate and objective control of exosome size and contamination after the purification process has also become critical. Based on professional knowledge and high-sensitivity equipment, Creative Proteomics is able to identify exosomes in a fast and cost-effective way. If you have a question or want general information, please feel free to contact us. We look forward to working on your next project.

References

1. Zlotogorski-Hurvitz, Ayelet, et al. "Human saliva-derived exosomes: comparing methods of isolation." Journal of Histochemistry & Cytochemistry 63.3 (2015): 181-189.
2. Kowal, Emma JK, et al. "Extracellular vesicle isolation and analysis by western blotting." Extracellular Vesicles. Humana Press, New York, NY, 2017. 143-152.

* For Research Use Only. Not for use in diagnostic procedures.