Hello Everyone,
I am trying to precipitate protein using organic solvents such as acetonitrile and isopropanol. The ppt occurs but as soon I add TFA (1%) or TFA in an organic solvent the ppt does not occur. Also, I tried using TCA up to 30% but no ppt. Does anyone have a similar issue when using TCA or any acid?
My objective is to ppt the protein and analyze the supernatant, which contains saponins by HPLC-UV.
Hey dude! I hope you are keeping well! We have had luck using saturated (NH4)2SO4 to ppt protein out. Add a small volume to the protein solution make it 5050 with MeCN and pellet out the protein.
Hi Paddy! I am doing well hope your are well too. Thanks for the suggestion. NH4SO4 won't evaporate that might give a disturbing signal with RPC-HPLC UV.
glad to hear you are well! We do this currently with enzymatic sample and run them on LC-MS (we clean the ion source regularly). you could also try spinning out your protein through a molecular weight cut off. i.e only the saponin would come through the membrane.
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