FBS interference in SDS PAGE - (May/01/2010 )
Hi everyone,
Fairly new to proteomics....I had cells that were grown in 10% FBS, washed them with a little PBS prior to harvesting and lysing. Now I have protein sup, but the sds-page gel looks horrible- can't help but think FBS is rearing its ugly head. Is there any way to wash it off while maintaining the integrity of all the other proteins?
nummy1987 on May 1 2010, 07:07 PM said:
Fairly new to proteomics....I had cells that were grown in 10% FBS, washed them with a little PBS prior to harvesting and lysing. Now I have protein sup, but the sds-page gel looks horrible- can't help but think FBS is rearing its ugly head. Is there any way to wash it off while maintaining the integrity of all the other proteins?
why with a little PBS? I would at least wash two times
I wanted to get the cells flash frozen ASAP so did one initial wash. Is there anyway now that it's protein sup to "wash" off FBS?
Inmost sun on May 1 2010, 04:30 PM said:
nummy1987 on May 1 2010, 07:07 PM said:
Fairly new to proteomics....I had cells that were grown in 10% FBS, washed them with a little PBS prior to harvesting and lysing. Now I have protein sup, but the sds-page gel looks horrible- can't help but think FBS is rearing its ugly head. Is there any way to wash it off while maintaining the integrity of all the other proteins?
why with a little PBS? I would at least wash two times
tere must be sum clean up kits for FBS... i m not suire though... morover if u are not able to remove the FBS.. try doing a western blot for ur protein of inerest... this way the blot wont look ugly too!!