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Double Bands in Western Blotting


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#1 Limp_chimp

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Posted 11 March 2004 - 07:39 AM

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!

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Edited by Limp_chimp, 12 March 2004 - 01:09 AM.


#2 keithwu

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Posted 14 March 2004 - 06:52 AM

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

#3 Limp_chimp

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Posted 15 March 2004 - 02:53 AM

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




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