ethanol precipitation of protein
Posted 27 July 2012 - 12:49 PM
I want to get information about ethanol precipitation method. I have produced a toxin from a fungal species in a volume of 2 liters and I want to purify the toxin by ion exchange chromatography. I will concentrate 50-fold the sample with ultrafiltration and then, I will apply buffer exchange for chromatography. But, because of total protein concentration of culture supernatant is very low (15 mg/2 liters), I am undecided whether or not the ethanol precipitation method cause a serious decrease in the amount of the total protein. Also, I dont want to lose the activity of the toxin. Do I see any problem about using this method? or Should I use centrifugal filters in this regard? I would appreciate if you share your experiences in this regard with me.Thank you
Posted 30 July 2012 - 07:10 AM
you can use ultrafiltration to concentrate and exchange buffer (diafiltration) then apply the sample to iec.
Edited by mdfenko, 30 July 2012 - 07:11 AM.
genius does what it must
i do what i get paid to do