TritonX-100 Lysis buffer recipe = 25 mM Tris•HCl pH 7.4, 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 5% glycerol
I am sure it won't break up nuclear membrane but not sure whether it can break up mitochondria membrane.
Thank you very much
If you sonicate cells in the buffer, I don't see a reason mitochondrial (membrane) proteins cannot be solubilized.
The buffer is strong enough for sure. If you do not want to sonicate then you may use 27 gauge syringe. You can also try let the cells sit in the buffer a longer period of time on ice. However, sonication maybe faster to achieve the lysis.
Mitochondria will be lysed in 1% Triton-X without any need for sonication, usually within a few minutes.
Yes it simply depends on the critical micelle concentration of the detergent used. In other words it depends on the ratio between mitochondrial membranes and detergent.
Hi Asa Wu,
1% TritonX will solublize mitochondrial membranes and release mitochondrial matrix proteins. The extent of this solublization depends on time and amount of lysis buffer for the cells/tissue. If you want a complete lysis of mitochindria, sonication can help or you can also use a stronger lysis buffer like RIPA.
Good luck :)
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