WB- proteins of lower-than-expected MW - (Aug/24/2005 )
I've been trying out westerns for some high molecular weight proteins (Tiam1- 177 kDa, and SRPK-92 kDa) and have been getting lower than expected bands by ECL. Tiam appears around 90kDa and SRPK comes around 60 kDa. I have used the same samples for some other proteins and i get the expected MW bands. My questions are the following:
a. could this be due to consistent degradation in all my samples? I prepared lysates a month ago and kept them stored in aliquots at -80 and -20 and have got these bands in both groups.
b. What other modifications could occur that would result in lower than expected MW bands??
Any suggestions would be highly appreciated...
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Well, I know this is one old topic, but I have now the same problem! My protein (SP1) has around 100kDa and i can`t find it in my WB's. Instead of this, I find a lower band. I made this question one month ago, but nobody could tell me what's happening. Did you find a solution? I though that there was a problem with some denaturing agent: temperature, detergent, ect.