Protocol Online logo
Top : New Forum Archives (2009-): : Protein and Proteomics

protein precipitation - (Apr/21/2009 )

I need to test whether my storage buffer will precipitate my protein. How can I know my protein is precipitated by the buffer?

-Nrelo-

centrifuge it and see if it develops a pellet (before and after changing the buffer).

-mdfenko-

Treat your sample with the storage buffer and a suitable control. If the protein precipitates, a pellet will be visible upon centrifugation. However, the pellet will be discernible only if the concentration of your protein is high enough to produce a large one. In case you do not see any pellet, run SDS-PAGE with the supernatant along with the control and comapre the intensity of the bands. If you have a functional assay for your protein then try it with the supernatant. Note: do not forget to inlcude a good control; this is a compatative anlaysis.

-sharath-

sharath on Apr 21 2009, 01:58 PM said:

Treat your sample with the storage buffer and a suitable control. If the protein precipitates, a pellet will be visible upon centrifugation. However, the pellet will be discernible only if the concentration of your protein is high enough to produce a large one. In case you do not see any pellet, run SDS-PAGE with the supernatant along with the control and comapre the intensity of the bands. If you have a functional assay for your protein then try it with the supernatant. Note: do not forget to inlcude a good control; this is a compatative anlaysis.


thx guys. My protein is an enzyme so I am going to use enzyme assay for testing

-Nrelo-