I have extracelluar vesicles isolated from serum and want to know which protein lysis buffer I should use? I usually use a standard RIPA buffer but have seen some people use a urea/SDS/leupeptin/pepstatin/PMSF mix. Which is best?
You can skip a lysis buffer and directly go for chloroform/methanol extraction which will also get rid of the lipids part of the EVs. Then pellet from this step can be resuspended in any buffer you like depending on your downstream application.
Hi there,
The nature of the buffer to use is dependent on the downstream experiments you intend to do with your protein sample: do you need native or denatured proteins? RIPA is OK for native protein extraction and the urea based buffer should be denaturant for proteins (depending on urea concentration).
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