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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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protein presipitation
Posted by: zia (IP Hidden, Unregistered user, )
Date: March 15, 2005 09:40AM
Respected,
If crude extract of protein (supernatent after filtration/centrifugation) is freez dried. How the salts or component of extration buffer can be removed from that protein? Thanks Zia
Re: protein presipitation
Posted by: bassamfahmawi (IP Hidden, Regular member, 42)
Date: March 16, 2005 07:47AM
Before freeze drying, you need to perform desalting of your samples by using desalting columns. Since you want to remove salts and extraction buffer dialyis would be the ideal choice, you can do this without ultering the concentration of your protein sample if you use a dialysis conctrating reagent (Check Pierce for this product at www.piercenet.com).
Good Luck!
Re: protein presipitation
Posted by: zia (IP Hidden, Unregistered user, )
Date: March 16, 2005 10:34AM
Respected,
its a good choice to use desalting columns or dialysis bag to remove salts or extraction buffer. one thing which i think and need yur comments is that if we precepitate the proteins by organic solvent and washing 2-3 times by same solvent, what are the chances that the salts may be present in that. is it possible? regards zia
Re: protein presipitation
Posted by: bassamfahmawi (IP Hidden, Regular member, 42)
Date: March 16, 2005 12:52PM
The use of organic solvents is a concentrating method of proteins and salt will also precipitate. In many cases the concentrated sample may not go back into solution and diffecult to reconstiute, therefoe the use of desalting and concentrating columns are ideal for your purpose.
Good Luck!
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