not "that" 2D gel - (Mar/28/2011 )
Hi,
I want to do a 2D gel analysis of a protein-substrate complex where the first dimension is SDS-PAGE and the second dimension Urea PAGE, can somebody direct me to correct literature/article or book chapter describing such a protocol. (alternatively, how do i get rid of the SDS from the SDS-PAGE gel slice after running the first dimension?)
Thanks,
New_to_biochem
-new_to_biochem-
personally, i would do the urea page first then the sds page.
sds can be removed by displacement with triton x-100 or nonidet p-40. however, you will probably lose a lot of the protein (or, at least, see it diffuse) if you perform this in-gel (which is why i would recommend performing sds-page last).
-mdfenko-
mdfenko on Tue Mar 29 16:30:58 2011 said:
personally, i would do the urea page first then the sds page.
sds can be removed by displacement with triton x-100 or nonidet p-40. however, you will probably lose a lot of the protein (or, at least, see it diffuse) if you perform this in-gel (which is why i would recommend performing sds-page last).
mdfenko, Thanks for your suggestion, but Urea-PAGE first and SDS next has been done and the information obtained from it insufficient for the complex. I will try triton x-100 or nonidet p-40 wash of the gel slice. Does anybody know if the old "Hoefer" 2D gel electrophoresis handbook is same as the new "GE" 2D gel ectrophoresis handbook?
Thanks in advance.
-new_to_biochem-