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stripping buffer for western blot


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3 replies to this topic

#1 farmtox

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Posted 04 May 2012 - 01:42 AM

hi!is there a mild stripping buffer for cerebellar substrates for western blot?

#2 memari

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Posted 05 May 2012 - 08:12 AM

https://encrypted.go...x-a&channel=rcs


Babak
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Babak Memari

#3 farmtox

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Posted 06 May 2012 - 01:37 AM

on that page there are different stripping buffers...in my lab,we use stripping buffer containing tris base,distilled water.pH 2 do you think it is too hard?

#4 memari

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Posted 06 May 2012 - 05:59 AM

http://www.abcam.com...20reprobing.pdf

Mild stripping
Buffer, 1 liter
15 g glycine
1 g SDS
10 ml Tween 20
Adjust pH to 2.2
Bring volume up to 1 L with ultrapure water.

Membrane incubation
Use a volume that will cover the membrane. Incubateat room temperature for 5-10 minutes.
Discard buffer.
5-10 minutes fresh stripping buffer.
Discard buffer.
10 minutes PBS
10 minutes PBS
5 minutes TBST
5 minutes TBST
Ready for blocking stage.
Harsh stripping
Prepare buffer and strip membranes under a fumehood.
Buffer, 0.1 litre
20 ml SDS 10%
12.5 ml Tris HCl pH 6.8 0.5 M

Edited by memari, 06 May 2012 - 06:00 AM.

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Babak Memari




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