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Result in the lab is not the same as published - (Nov/03/2014 )

Hi,

Iam working with a meschencymal cell line. My protein of interest is around 57 kDa and there is a published report in this cell line for the same protein. But the problem is, when I do the same western blot, and I use the same antibody as they used in the report, Iam not able to get it band in the same size and my size is always less than 50. Can anyone explain this behavior of the cell line. what are the possibility for getting this

-jaya2020-

There are a number of possibilities:

 

Different type of gel (e.g. native PAGE vs SDS-PAGE, Bis-tris vs tris-tricine) being run?

Antibody binding non-specifically (of either you or the paper)?

Incomplete denaturation in the paper?

Incorrect labeling of figure in the paper/typographical error?

They were using a transfected-tagged protein?

One of you is working with the wrong cell line (contaminated? mis-labeled?)

-bob1-

could also be the result of proteolysis. they may be working faster or colder, may have forgotten to mention inhibitors. your inhibitors may be past their useful date.

 

if sds-page, one of you may have sample buffer past its prime, improper heating of the sample. if you pour your own gels (or they do), it may not be completely polymerized (aged) when run.

 

the list can go on and on....

-mdfenko-

Thanks. I will try to eliminate the possibilites as far as possible

-jaya2020-