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Problem with Immunoprecipitated samples


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#1 bochum

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Posted 14 September 2011 - 04:37 AM

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 12% SDS-polyacrylamide gel?
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Martin
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#2 bob1

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Posted 14 September 2011 - 10:00 PM

Have you run this protein out on a gel before? If so, does it run at the expected size?

If not - have a look in the literature for your protein; some proteins do not run as their molecular mass would predict.

#3 deadally

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Posted 27 September 2011 - 02:40 AM

Exactly like Bob said, you should check the literature, since permanent PTMs like glycosylation can have a massive effect on the apparent molecular weight (see CD44 for a good example)




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