Protocol Online logo
Top : New Forum Archives (2009-): : Protein and Proteomics

nonspecific band in transfected c-abl cells - nonspecific band in transfected c-abl cells (Jan/14/2010 )

hi everyone,

I am trying to see knock-down of c-abl gene in my siRNA transfected cells by real-time and western blot.
while, i got knock-down levels 80% by real-time and my western also show a 50%reduction in estimated band size 120Kd compared to control.
My problem is that i am getting a smaller band around 35kda in transfected samples but not in controls.
I have no idea what is this band? Is it fragmented part of my protein in my transfected samples because of RNAi effect which is not there in controls.If it is, how can i prove it? Is there any way to find out so. Any suggestions are welcome to solve it.



You can cut out the band from a gel and "sequence" it using mass spectrometry - have a talk to a biochem department, they should be able to help.