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Optimal protocols for restripping membranes

Optimal protocols for restripping membranes - Western Blot Forum

Optimal protocols for restripping membranes - 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 06-21-2008, 04:53 AM
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Default Optimal protocols for restripping membranes



Hey I've got a lot of membranes to reprobe and when I've done it in the past its been quite inconsistent in terms of the quality of the second exposures. Is there anyone out there that may have some pointers about protocols to keep the signal clear and the background low. We have some very limited (therefore) precious samples and need to do many strips.

Anthony
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Old 06-21-2008, 03:13 PM
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Default Re: Optimal protocols for restripping membranes

I used Restore Plus (Pierce) ---> [Only registered users see links. ]

Usually it takes 15 minutes (but some Ab bind very strong and will require two 15 minute incubations).
Another tip is to optimize Antibody concentrations, the less Ab the more effective the stripping process.
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Old 06-23-2008, 01:01 AM
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Default Re: Optimal protocols for restripping membranes

danfive, what is the difference between the product you recommend and a stripping buffer of beta-mercaptoethanol, SDS, Tris HCL pH=6.8. Do you find from your experience that Restore Plus works better?

Last edited by agador; 06-23-2008 at 01:12 AM.
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Old 06-23-2008, 05:13 PM
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Post Re: Optimal protocols for restripping membranes

Quote:
Originally Posted by agador View Post
danfive, what is the difference between the product you recommend and a stripping buffer of beta-mercaptoethanol, SDS, Tris HCL pH=6.8. Do you find from your experience that Restore Plus works better?
The difference will be that this product won't smell like beta-merc, and probably won't need as much washes like SDS. Those are obvious matters of convenience and not performance. You can definitely make your own.
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Old 06-24-2008, 08:57 PM
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Default Re: Optimal protocols for restripping membranes

danfive,
Can you reuse Restore Plus after stripping a membrane or do you have to discard it after each use... it might be expensive after a while
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Old 06-24-2008, 09:16 PM
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Default Re: Optimal protocols for restripping membranes

Discard it.
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