We did ultracentrifugation to purify exosomes, but we only obtain 4 ug of protein (BCA assay of intact exosomes) from 2 mL of plasma. This is a very low yield. We need at least 40 ug to do flow cytometry characterization and 10 ug to do TEM characterization. Can someone recommend me a protocol to obtain this amount of exosomes? Should I homogenize the exosomes before protein determination?
I would recommend using the protocol, described in this article:
Optimized exosome isolation protocol for cell culture supernatant and human plasma
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507751/
Exo-spin Kit is a good choice.
An alternative would be the detection and separation by FACS, or even direct detection in plasma by flow cytometry with a simple pretreatment and with very small amount of sample. I might recommend you to try a detection kit extremely sensitive (LOD
Wu, C. X., & Liu, Z. F. (2018). Proteomic profiling of sweat exosome suggests its involvement in skin immunity. Journal of Investigative Dermatology, 138(1), 89-97.
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