Hello
Does NP-40 or Triton X-100 lysis buffer lyse/break mitochondrial and nuclear membranes to release out their proteins?
In addition, RIPA can rip off all organelle membranes and RIPA = NP-40 or Triton X-100 lysis buffer + SDS + sodium deoxycholate. Can RIPA without SDS still lyse all membrane? From what I read, SDS seem play role only in denaturing protein only, not lyse cell.
Thank you
I think NP-40 or Triton-X-100 with proper concentration is sufficient to lyse cells/cell membranes including cell organelles.
RIPA is more denaturing than NP40 buffer alone and should not be used for fragile enzymes.
NP40 buffer is a name for RIPA lacking sodium deoxycholate and SDS. This lysis buffer preserves kinase activity better than RIPA, but gives a higher background in the immunoprecipitate. NP40 buffer can be made with either NP40 or Triton X-100, and can be buffered with either phosphate or Tris, depending on your needs.
You can use 0.1% Triton-X100 in order to lyse the membrane mtitohondry. An activity of many enzymes, including succinate dehydrogenase associared with the membrane will be saved.
Excellent review paper on the effects of Triton X on mitochondrial proteins.
Triton X-100 solubilization of mitochondrial inner and outer membranes
J. I. G. Gurtubay DOI 10.1007/BF00745012
Dodecyl-beta-D-maltoside is another detergent you might consider...depends on what proteins you are hoping to isolate - whether membrane bound or not
This paper might help you. It discusses method of choice for disruption of cells prior to mitochondrial isolation.
http://www.sciencedirect.com/science/article/pii/S0003269709005004
Maritza Kruger Can RIPA work for high molecular weight enzyme which is a nuclear protein?
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