Protocol Online logo
Top : New Forum Archives (2009-): : SDS-PAGE and Western Blotting

SDS-PAGE/2D gel and western blot for multiple proteins for multiple sample - (Jul/05/2018 )

Respected members and Researchers,

  i am doing my experiment on SDS PAGE and 2D gel electrophoresis. Further I have to do the confirmation of my protein by western blot. In one SDS gel there are 10  samples and every lane have 4 proteins in target and my sample is about more than 100. So to perform western blot for all 4 protein for different lanes is not an easy task, As I wants expert opinion from your experiences, what would you suggest? Do I cut all the 4 proteins of my interest from every lane or can I perform all lanes together at least one gel having 10 lanes at 4 successive steps for 4 different proteins?

 

Thank You very much

-Faisal SHAH-

you may be able to detect all 4 proteins at the same time. this depends on how well separated the proteins and how specific the antibodies are. also, the source of the antibodies (in which species they are made) for selecting secondary antibody.

-mdfenko-