Hi all,
I've been using the Exo-Quick reagent by SBI for extracting exosomes from cultured medium from MSCs. The MSCs are cultured in exosome-free FBS for 3D prior to exosome extraction. The exosomes are diluted in Ripa, protein quantified via BCA and ran on WBs (25-50ug). In my attempts to label them with different markers, i keep seeing a consistent 38kDa (and maybe one at 31kDa too) band regardless of the species of the primary/secondary used and the marker used.. I've tried CD63, 9, 81, TSG101, Actinin4, Flotilin, Mitofilin etc. Most of the times, I have failed to get the actual band too - nothing for any of the above markers except for Flotilin. I've tried both reducing and non-reducing conditions. Most the the antibodies are giving bands in P4 pellets from other cells (colleague's work) so the are most likely not the issue.
Any suggestions at all please?
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