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Stripping the membrane for LiCor


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#1 LacquerHead

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Posted 18 November 2012 - 08:59 AM

Hi all,

I am wondering if anyone has found good conditions and stripping buffer formula for stripping the membrane to re-probe on LiCor? I find that when I strip my PVDF membrane its much harder to do so for LiCor than film and there is more background. Any suggestions here would be great thanks!

The stripping buffer I am currently using is:

20ml SDS 10%
12.5ml Tris HCl 0.5M pH 6.8
800ul BME
67.5ml dH20

Leaving it at 50 degrees for 45 minutes before extensive washes with water and TBST and reblocking.

#2 bob1

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Posted 18 November 2012 - 11:34 AM

PVDF has a higher autofluorescence than nitrocellulose which is why you have the background. The fluorescence function is also more sensitive to background than film is. Your stripping protocol should work fine, but you will need to re-block your membranes and treat them as you would normally.

If you want to do this well, make sure that you are using a fluorescence rated PVDF or switch to nitrocellulose.




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