Trying to prepare cytosolic and nuclear protein fractions of S.cerevisiae. I am using anti-nsp1 antibody (against yeast nucleoporin, Nsp1) as a marker of the nuclear protein fraction. It gives an expected size band (about 100Kd) with the total yeast lysate and instead of the 100Kd band, gives a (approx.) 50Kd doublet in the subcellular fractions.
The antibody is supposed to be monoclonal.
Has anyone observed this doublet before? Are these 2 bands two domains / halves of Nsp1? Or are they simply non-specific bands that cross react with this antibody? Any input will be welcome.
Thanks!
Anupama Shanmuganathan
Graduate Student
Georgia State Univ., Atlanta, GA
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