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-   -   Why the band at wrong molecular weight? (http://www.molecularstation.com/forum/western-blot-forum/65398-why-band-wrong-molecular-weight.html)

Gorbin 03-30-2009 07:41 AM

Why the band at wrong molecular weight?
 
The single band detected by WB is about 50kDa smaller than it should be. What are the possible reasons other than complete degradation?

admin 03-30-2009 11:35 AM

Re: Why the band at wrong molecular weight?
 
Hi Gorbin,
how big is your protein? Also 50kDa is quite a bit of a difference, could be proteosomal degradation, splicing, or other like a gel problem or ladder issue.

Have you checked your protein sequence / protein published information to see if it behaves funny?

Gorbin 03-31-2009 09:47 AM

Re: Why the band at wrong molecular weight?
 
Thank you for your kind suggestion.
The protein here is actually HIF1-alpha, which is extremely vulnerable to degradation. In literature it appears a band at around 120 kDa. But you know, most people won't show the entire gel, so I have no idea if they also have a 70kDa band. It is possible that the 70kDa band is the degraded form of HIF1-alpha. Is there any way to check it, rather than blindly taking more care of prevention of protein degradation? Thanks!


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