No peak for gel filtration for soluble protein - Need help (Mar/07/2005 )
I purified a MBP-tagged protein, which is quited soluble. On SDS-PAGE, the MW is about 65 KDa and it's a very thick band. However, when I run the protein elution on gel filtration, both 75 and 200 column, there is no any peak for protein. The running buffer I used is the same as elution buffer for purification.
I really don't know what happened? Did my protein degraded or it's stucked on the column?
Thanks for your help! I am really frustrated about this.
Are you confident in the loading procedure of gel filtration?
in my lab, some labmate load the protein wrongly and result in loss of all protein