I use cytoplasmatic and nuclear extraction protocol for my cell. Normally the cell pellet (for example, Hela, 2FTGH, U3A, U6A) will dissolve in both extraction buffer.
However,Jurkat and Romos cell pellet (they are suspension cell line) do not dissovle in nuclear extraction buffer after centrifugation of adding cytoplasmatic extraction buffer.
They are too sticky enough even I tried to pipet up and down or vortex them.
Do anyone of you face this problem before? Any solution?
One idea is to be more gentle when lysing your cells. It is possible that you are lysing both the plasma and nuclear membrane when pipetting/vortexing. The contents of the nucleus can become quite sticky to work with.
Hi Khalid,
I do the same thing to other cells as well. They work fine (pellet dissolve well in nuclear extraction buffer). However, it does not work for Jurkat and Ramos cell
One thing that you might try is to substitute your nuclear extraction buffer with a 3 detergent RIPA. Also instead of vortexing, it may be a good idea to manually disrupt the pellet with a p1000. After your protocol is finished, sonicate and that should completely disrupt any remaining stickiness
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