Hi,
We want to investigate the extracellular vesicle proteome through proteomics. Reading the literature in this regard, we are not clear about the necessary micrograms of extracellular vesicles for the realization of this approach.
Thank you in advance for any responses we may receive on this.
Hi Antonio,
the amount of extracellular vesicles (EV) required for a proteomic approach is dependent on the protein content of the EVs of interest which could differ in different sources. I would suggest that you perform a protein analysis from you EV preparation and adjust the amount that you need for your proteomics approach from this protein analysis. In one of our recent publications, Cilibrasi et al. 2022 "Definition of an Inflammatory Biomarker Signature in Plasma-Derived Extracellular Vesicles of Glioblastoma Patients" we have used 100ng of proteins from EVs to perform an SDS PAGE and do InGel digestion of 5 slices from each lane to get tryptic peptides for downstream LC-MS analysis.
This amount worked good for us but the required amount can be very different in relation to the method and LC-MS equipment you are planning to use.
I hope this helps.
All the best,
Murat