I have taking the cell lysate and adding DSS which cross links primary amines. Since I haven't purified ASC, what are the odds that I would find ASC getting cross linked to other non-asc proteins since all proteins have an N-terminal primary amines and lysines have this group too? I haven't seen this issue being addressed in any of the protocols.
Hello Monyer,
I referred to the same protocol but it doesn't address the problem I mentioned in the question. Since the protein isn't being purified, the mixture may have non-specific protein cross-linking and even if one ASC cross-links with any random protein (s), One would still see that as a band in Western at a different molecular weight even though it is not a an ASC oligomer.
Hi
i think that I referred to the same protocol but it doesn't address the problem I mentioned in the question. Since the protein isn't being purified, the mixture may have non-specific protein cross-linking and even if one ASC cross-links with any random protein (s), One would still see that as a band in Western at a different molecular weight even though it is not a an ASC oligomer.
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