not for isolation i think oly protocol for analysis and purification... (such as, HPLC, MS, )
If you want to isolate a specific protein then immunoprecipitation is the choice......
You can use RIPA-buffer or CST-buffer (Cell signalling technologies)
http://www.cellsignal.com/products/9806.html
http://www.cellsignal.com/products/9803.html
It's also possible to lysate your cells in Sample buffer for SDS-PAGE. But than you can't measure protein concentration. In this case you can use house keeping genes (HSP90, ß-actin) for normalization.
Hi,
I would like to use the protein extract as an antigen for Dendritic cells. Is there anyway to neutralize the lysis buffer, so that it doesnt affect my dendritic cells?
thank you
Most of the lysis buffers composes detergents (to remove membrane bound proteins) which has to be removed by Dialysis or precipitation methods. For using whole cell extract as an antigen for Dendritic cell, you can do lysis using mild sonication or any physical disruption methods(Freeze thaw/Syringe lysis) in PBS Buffer only.
Thanks for your response Chandra. I already tried the freeze thaw method, but unfortunately that was toxic to the dendritic cells. Could you tell me a little bit more about the syringe lysis method or point me to a protocol for it?
Thanks again.
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