For purification of this protein i used lysis buffer( 20mM tri s HCl pH=8,with 0.5M Nacl and 0.2M imidazol), (20mM NaPB pH=6.5,with 0.5M Nacl and and 0.2M imidazol), (1x PBSl pH=7,and 0.5M imidazol).But I observed protein precipitation in all case.
If the protein was already insoluble before you lysed the cells, then it doesn't matter what lysis buffer you use, you will always get insoluble protein.
If the protein was at least somewhat soluble but is insoluble at high salt concentration, then you might improve the recovery of soluble protein by using a lower salt lysis buffer, since all of yours are quite salty.
Dear Adam, Thank you for your answer. I got the soluble protein after elute of resin with 20mM tris HCl pH=8,with 0.5M Nacl and 0.2M imidazol. I also thought like you that may be high salt conc. is a problem. Thats why I reduce the salt conc. from 0.5 M to 0.2 M but in that case i got protein precipitation again.
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