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stripping buffer

stripping buffer - Western Blot Forum

stripping buffer - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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  #1  
Old 07-13-2010, 07:41 PM
Pipette Filler
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Default stripping buffer



I need to make stripping buffer. The protocol that I have from GE says to use 100uM 2-mercaptoethanol, but the bottle I have from ProPure does not have molarity on the label. I went to the website and could not find any info sheets about the product. Does anyone have suggestions, or another stripping buffer protocol. Thanks.
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Old 07-19-2010, 01:45 PM
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Default Re: stripping buffer

Here is the composition of my stripping solution, which works very well for me:

- Beta mercapto ethanol (Sigma Cat.# M7154) = 700 l
- SDS 10% = 20 ml
- Tris HCl 0.5 M (pH 6.8) = 12.5 ml
- NaCl 5 M = 10 ml
- Complete to 100 ml with Water

Incubate your membranes in this solution heated at 50C for 30 min under gentle agitation (Care: This has to be done in a fume hood).

After the stripping, wash your membranes for at least 1 hour with your usual washing solution (change the solution 3-4 times).

Your membranes are ready to be re-probed.

Natty
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Old 07-19-2010, 10:16 PM
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Default Re: stripping buffer

Hi Natty,

Thanks for the information. This is very helpful.

-Wolfman
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Old 07-30-2010, 03:54 AM
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Default Re: stripping buffer

very helpful!
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