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urea/glycerol PAGE and cutting out the band... - cant find out protocols (Apr/24/2006 )

Dear friends!

so I have this problem i'm breaking my head over, I just can't find the proper protocols for my purpose (s).....Breifly (for ones who dont already know smile.gif) I have expressed protein from E coli which has some degraded bands on SDS-PAGE...I want to isolate the upper band....since my protein is insoluble rolleyes.gif ...i have prepared it as inclusion body in 8M urea....now i want to use urea PAGE (with glycerol) to isolate the band and cut it out from the gel....then dialyse it.... blink.gif ....my professor gave me some table of urea/glycerol PAGe buffers but it is so not clear and not complete.... sad.gif ....and i cant find out any protocol to tell me how actually I dissolve this band out of the gel?!?! I am still searching and maybe will find out some protocols....will appreciate any help so so much....thank you!

-Kathy-

unsure.gif ...anyone?? and if possible do you think voltage settings should vary or same as SDS-PAGE?? ....

-Kathy-