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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-

nummy1987 on May 1 2010, 07:07 PM said:

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?

why with a little PBS? I would at least wash two times

-Inmost sun-

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:

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?

why with a little PBS? I would at least wash two times

-nummy1987-

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!!

-Prep!-