TCA precipitation of proteins - (Jan/26/2004 )

Try a 1:1 dilution of the recovered fraction using buffer containing no Triton

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Hello,

I am trying to isolate a protein who has a cysteine rersidue. I am testing if this cysteine is making a disulfide bond with another monomer as a mode of regulation. I found a paper who was saying that proetins redox state can be fixed by pption with TCA but it doe snot give a detailed protocol plus they work on E.c oli while i am working on bacillus. so anyone could help me what to do exactly? Thank you in advance.

ali

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Anybody knows if TCA precipitation affect to protein phosphorilation?

Thanks a lot

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seems not there is a EMBO Paper adressing this Delaunay A et al "H2O2 sensing through oxidation of the Yap1 transcription factor" see Meterials and methods!
does anyone did an IP after a TCA precipitation?

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Hello,

I'm trying to apply TCA precipitation in the lab, but I couldn't dissolve the pellet in Laemli's buffer with DTT. Does anyone experience the same problem? And how did oyu overcome?

By the way, it is very important for me to dissolve this pellet in lysis buffer, because I have to quantify before loading.

Thanks a lot

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how do you try to dissolve?
i use 37° heating with agitation for up to 1H...

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