Immunoprecipitated samples

[pmitra]

A protein that I immunoprecipitated from cell lysate, runs a little bit higher than the expected size. Has any one observed IP samples running at incorrect molecular weights on a 10% SDS-polyacrylamide gel?

[obama]

it will easy to answer, if you mention what was the expected size of your band and where you getting the band.

[pmitra]

Thanks for the quick response. The expected size is ~95 kDa; the immunoprecipitated band elutes between the 95 kDa and 130 kDa bands of the protein ladder.

Thanks!

[obama]

what type of protein is that, if it is glycoprotein. some time after immunoprecipitation and other process leads to some changes in such protein. there by changing it charge or some even MW of such protein because (carbohydrate moity in it). I think this shift may be because of that...

[pmitra]

Thanks for your input!