Double Bands in Western Blotting - Please Help (Mar/11/2004 )

Hi Everyone,
I have been assessing the expression of Angiotensin II, AT1 receptor proteins in rat synovial tissue. When analysing my gels, i find that there are two bands in the region of interest, situated very close to each other. I realise this could be due to phosphorylation or oxidation, however i am unsure of how to get rid of this problem.

Is this a serious problem that could prevent publication?

How could i assess whether or not this is due to phosphorylation or oxidisation?

I have attached a recent picture...i have cleaner ones, but not on this computer!

hi

try incubate your protein sample with phosphotase which dephosphorate most phosphoprotein.

once you confirmed the extra band is the phospho form, I think it would not affect your publication

Excellent...thanx for the suggestion, i shall give it a go!!