In WB FANCD2 has 2 bands (L-FANCD2 and S-FANCD2) normally seen as ~150k dalton bands. In immunoblotting of FANCD2 with (X)-protein FANCD2 band appeared at 100k dalton band. What are the possible explanations for this band shift?
Not sure what you mean by immunoblotting FANCD2 with (X)-protein... But three things come to mind, (1) the band you're seeing is not FANCD2, (2) FANCD2 is being proteolytically cleaved, or (3) there is a splice mutant that is generating a shorter form. If this is over expressed protein off of a plasmid, you can probably rule out the third option. To rule out the first option, you can knock down FANCD2 with siRNA/shRNA or knock it out with CRISPR. If the band goes away, then it is very likely FANCD2. You could also try another FANCD2 antibody, but if your lower molecular weight band is missing the epitope, let's say through cleavage, then you'll not see it. Good luck!
Hello
I agree with Michael 100% , you can see this link may help you
Article FANCI protein binds to DNA and interacts with FANCD2 to reco...
Good Luck
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