Hello, I am doing a western blot using crude extract of leismania for the detection of a basic protein by western blot. The expected size is 17.5 but on my western blot i have a very good band but with a MW of about 14.7.
Any ideas?
Thank you very much
I agree that some proteins do not run true to size on SDS_PAGE. The thing I would worry about is that your protein has been cleaved and may be missing one end. If it is not glycosylated, you could check by mass spec.
Thank you very much for the answers. I always keep my samples on Ice and my Lysis buffer has proteases inhibitor. Before the loading I heat the samples with the loading buffer at 95 C for 7 min.
I was wondering if the faster migration could be related to the differences of the ratio between SDS per mass of protein of my protein on comparison to the ladder, Is it possible that basic protein (PI= 10.17) could take more SDS than more neutral proteins? That meaning more negative charge per mas of protein and more negative charge so faster migration?
Best regards
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