Hi everyone, I'm wondering if someone could help me. I want to evaluate the presence of specific proteins in the interior of exosomes but can't find any protocol that would allow us to permeabilize them for use in ELISA (R&D). My question is, if someone knows if the assay diluents, usually present in the ELISA kits, have any detergent in its composition that may act as a permeabilizer for my exosomes.
Thank you all in advance.
Maybe this could help:
first exclude if the proteins you are interested in are on the outside of the EVs (i.e. by a bead coupled FACS, so without detergents breaking your membranes). So; coat beads with protein A/G with antibody your interested in, and as a second antibody use CD81/63/9 etc.
if it is not, you can do ELISA's or westernblots with the iosolated EVs (so breaking membranes with i,e RIPA buffer with SDS )
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA