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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? |
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? |
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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