Why wet blot cause non-specific signal

Alberto mrealpe at ibt.unam.mx
Tue Jan 2 12:56:14 EST 2001


Remember, most of detection systems are immunological in nature, although
antisera are used after get wet again it is not the same fresh proteins
than rewet ones.  I think the matter is a quality one, but not ruling out
a decrease in quantity as well.  To answer your question, wet the blot
is getting again native conformation to your blotted proteins however they
never will be as before getting dry (at least in a certain degree).

Check by densitometry the staining degree after dry (ponceau stain), I
guess after several cycles of drying and wetting the quantity should be
severely affected (consider every wash in a general western procedure).

Hope this could help.  Luck !!.

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	MAURICIO REALPE
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On 2 Jan 2001, Kittapong Tang wrote:

> Hi All,
>    I have some problems in protein botting. Why wet blot give more signal 
> than semi-dry blot even with the same sample and the same blotting 
> efficiency? Can wet blot change the protein conformation?
> 
> Thanks in advance for all help.
> 
> Tang K.
> _________________________________________________________________________
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