Hi, I have been isolating exosomes from HUVECs culture and wanted to run a western blot for the characterization of exosomes. Till date, I have only checked for CD63 marker and it has worked. Since I plan to expose the HUVECs to high glucose conditions in the future, I was wondering if there are any other exosome markers that doesn't get affected by these conditions?
I read that exosome markers CD81 and HSP70 expressions change under diabetic conditions, so I cannot use those.
Any suggestions would be appreciated. Thanks
If the method of detection of markers is Western blotting, which is semi-quantitative, I would check for all exosomal protein markers (including the CD81 and HSP70). It is likely that, depending upon the sensitivity of detection, you might see some minor changes in the levels of these (and other) markers.
Cell signaling has an Exo marker sampler kit
https://www.cellsignal.com/products/primary-antibodies/exosomal-marker-antibody-sampler-kit/74220
to test various markers and see which work best for your cells and conditions
also you can look at SBI's dot blot membranes for which markers to choose, these are not very quantitative though, but you can choose an Ab based on it.
https://systembio.com/shop/exo-check-exosome-antibody-array/
also check the ISEV 2018 minimum guidelines paper, to verify exosomes you need 3 positive and 1 negative marker, and all should not be tetraspanins, so you don't get dinged by reviewers
Article Journal of Extracellular Vesicles Minimal information for st...
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