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protein precipitation


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3 replies to this topic

#1 Nrelo

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Posted 21 April 2009 - 02:17 AM

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

#2 mdfenko

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Posted 21 April 2009 - 08:13 AM

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

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Posted 21 April 2009 - 01:58 PM

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 B.

#4 Nrelo

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Posted 22 April 2009 - 01:52 AM

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




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