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can stripping buffer be re-used?

can stripping buffer be re-used? - Western Blot Forum

can stripping buffer be re-used? - 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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Old 07-21-2008, 07:41 PM
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Default can stripping buffer be re-used?



my stripping buffer works great for the first time , but when I reuse it ,

seems it doesn't work , can you guys who have ever re-used it ?
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Old 07-22-2008, 06:14 AM
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Default Re: can stripping buffer be re-used?

Hello Lynch, i know some people that have reused it however would be good to know what doesnt work: is it the stripping of the antibodies or later exposure?

cheers
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Old 07-22-2008, 05:32 PM
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Default Re: can stripping buffer be re-used?

The common homemade stripping buffers contain 2-ME to break the dissulfide bonds on the IgG and help remove them. 2-ME is going to evaporate more quickly during a 60C stripping incubation. The lower amount of 2-ME is going to reduce your stripping efficiency. Success on reuse is going to depend on how much effort is required to get those IgG to let go. If you are stripping an actin band, I would use fresh to make sure you don't have to repeat the whole thing again. Saving a little money now can waste a lot of time later.
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Old 07-23-2008, 04:24 AM
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Default Re: can stripping buffer be re-used?

thanks Kevin great response. Kevin is right, if your stripping buffer does not smell then probably most of the 2-ME is gone.
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Old 07-23-2008, 01:23 PM
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Default Re: can stripping buffer be re-used?

Thanks , kevin, I finally know the principle of stripping buffer, it's really helpful.
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