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Stripping Solution

Stripping Solution - Protocols and Methods Forum

Stripping Solution - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 10-15-2006, 07:02 PM
Maryam
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Default Stripping Solution



Does anybody know if NaOH solution can be used as a stripping solution? if So, what should be the final concentration?
Thanks


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  #2  
Old 10-15-2006, 11:15 PM
Christian Praetorius
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Default Stripping Solution

Maryam <[Only registered users see links. ]> wrote:


What do you want to strip?

Christian

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  #3  
Old 10-16-2006, 06:54 PM
WS
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Default Stripping Solution

Dear Maryam,

I assume you want to strip westerns.

0.2 M NaOH in water might do the trick. There are commercial
preparations (eg Chemicon ReBlot Plus) which come as 10% concentrate.
They are alkaline and the MSDS says 2.4N NaOH (light) and 3.6M (strong)
also containing 0.05% sodium azide. The latter is intended to kill the
peroxidase from the secondary AB.

You might shortly rinse the membrane before and after with water in
order to limit carryover of blocking buffer / ecl resp. stripping
solution.

In my experience, a 15 min incubation in one of these solutions
sometimes, but not always does a good job. Do not expect that
previously strong signals will disappear completely. in order to check
your success, do a reblot with secondary AB only.

Good luck,

Wolfgang

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  #4  
Old 10-23-2006, 09:20 PM
Pow Joshi
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Default Stripping Solution

On 10/15/06, Christian Praetorius <[Only registered users see links. ]> wrote:

you might want to add some beta mercaptoethanol or DTT as well ...
this would cleave the -S-S- linkages in the antibodies (again assuming
you want to strip western blots)

pow

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  #5  
Old 10-26-2006, 02:59 PM
Pow Joshi
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Default Stripping Solution

On 10/24/06, [Only registered users see links. ] <[Only registered users see links. ]> wrote:

Dear Wolfgang,

frankly, I have used the stripping protocol aboout 3 years ago. I did
try to find it in my old notebooks last night, however without any
luck. I shall get back to you as soon as I lay my hands on it ... I
did check other protocols and they use 100mM beta-mercaptoethanol.

hope this would help,
pow



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