Dear All,
I want lyse whole (including nucleus) SY5Y cell without mechanical disrupting for Co-IP.
I have read many resources, and it can be done by either adding 0.1-0.25% Sodium Deoxycholate or increase NaCl to 420mM, in TritonX100 lysis buffer (50 mM Tris•HCl, pH 8.5, 150 mM NaCl, 1%TritonX100).
Which one is better to lyse whole cell for Co-IP?
Thank you
Cheerios,
Results for me were better with the second set (NaCl, Triton X-100). Hope that works for you too.
I got nice results when using 50 mM tris pH 8.0, 150 mM NaCl, 0.25% Sodium Deoxycholate, 1% Tritón X-100 or NP40, 0.1% SDS and 1 mM EDTA. My protein is nuclear and this was the only way to solubilize it with a good yield so as to perform Co-IP with the whole cell lysate. Hope it will help you.
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