Need to increase protein precipitation yield w/ TCA precipitation protocol - (Jun/01/2007 )

Hi all,

Whenever I do TCA precipitation w/ the eluted protein solution after immunoprecipitation, I always feel that the precipitated protein yield is too low. Is there any way to increase protein yield with TCA precipitation?

Great thanks.

Add first acetone and then TCA. normally it's 30' at -20°.
2h at -80° or more increase the yield.
You may also try lyophilisation for getting higher yield. But often you need dialysis after that as the salts are also lyophilized

if you are not going to apply to a gel and silver stain then you can add a few micrograms of trna to the solution before you add the tca. it will coprecipitate and carry the protein down when you spin it. we used to use this to concentrate samples for coomassie stained gels (we found that we would get a big orange blotch if we silver stained).