Protein dissolved - After acetone precipitation (Jul/27/2011 )
I am doning acetone precipitation of protease enzyme from culture medium. After precipitated, the pellet was resuspended with 0.01 M phosphate buffer pH 6.8. The protein sample dissolved but It form to mucilage. the sample was dialysed wiht the same buffer but it didn't change. Then, I used membrane cut off (50 kDa). the enzyme activity be lost. I wana to reform mucilage protein to solution for ionexchange step. this sample couldn't pass a column. How can I doing wiht this sample ?
thanks for any suggestions or help.
was the buffer just phosphate? if so then some protein may have bound to the membrane. ionic strength should be around 0.2M to prevent binding.
your protein may have been too concentrated or it may have aggregated. some of the proteases we work with readily aggregate.
When looking to start using protein supplements, there's a few things you have to determine before just going ahead and using them. The first thing you should be looking at is what your purpose for using supplements are.iPhone