I isolated exosomes from human plasma and would like to test the presence of certain proteins in the exosomes. Which is the best way of lysing exosomes before doing the ELIsa? Is it enough to resuspend them in equal volume of RIPA buffer (now they are in PBS)? Thanks!
SDS may effect the protein-Ab binding ......Try Sonication or and in combination with hypotonic solution....I never try this but should work in my opinion..Good luck.
https://www.researchgate.net/post/Is_it_possible_to_use_protein_extraction_with_RIPA_buffer_to_measure_kinase_activity_by_ELISA2
Hi. I assume that your ELISA plate is coated with specific antibody against a protein you are looking into the exosomes.
After adding 100 uL of exosome sample in ELISA wells, simply add 25 uL of disruption buffer (0.05% Tween20, 2.5% Triton X100, 0.02% thimerosal (optional) in PBS, pH 7.4). Incubate for 2 h at 37oC.
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