I am going to use T-PER tissue extraction (Thermo-Fisher) buffer with Roche ultra Protease and Phostop to extract phospho-protein from mouse spleen and liver. I will collect fresh tissue in buffer mentioned above and make homogenates to extract protein. However due to over load of subsequent other works on that day, I am thinking to freeze whole tissue lysate on that day. And will centrifuge and isolate protein on next available day. But I am concerning, if the salts in buffer and inhibitors may interfare phospho-proteins during freeze, thaw and later application. Does anybody stored tissue-protein lysate in lysis buffer? Or does anybody had suggestions in this regard or methods to isolate better quantities of phospho-proteins?
Hi Saraswoti,
I have worked with knock-out mice brain tissues way back in 2003 studying the metalloprotiens' trafficking in these tissues via Northern Blots. Those days, I have stored them in their lysis buffers for fututre work and it has worked for me.
Good luck.
It should be OK if you add protease inhibitor in you lysis buffer in it. It can be stored at -80 for several months at least if you are working with phopohrylated protein.
Good luck with you experiment!
you will be fine. The real issue occurs when your samples go through multiple freeze thaws. As long as you have multiple aliquots and store at -80, your samples will be fine
i agree with kartyk, they work just fine
best is if you aliquot them
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