Re: Whole mount Immunos

From:Jill McVee <> (by way of histonet)

>Date: Wed, 6 Sep 2000 12:56:31 +0100
>To: Jill McVee <>
>From: N Kenneison <>
>Subject: Re: Whole mount Immunos
>X-Mailer: Turnpike (32) Version 3.05 <fLNX3a+t8ftnba84n3FvrffHCE>
>hi there Jill
>I must confesss that I have never heard of the back and forth to buffer
>technique before - and I do only work with human tissue.
>However the usual cause for background is either the antibody is either
>over dilute or under dilute it may also be that the secondary antibody
>is binding with something - that is if you are using a secondary ab,
>also is it a multispecies link if so just use a specific secondary and
>not a multi link this may work.
>In message <>, Jill
>McVee <> writes
>>Dear Histonetters,
>>                  I am currently engaged in staining whole mount frog
>>embryo nervous systems for immuno and although I,ve done the usual
>>things(diluted AB, blocked for endogenous peroxidase, added things to the
>>Normal serum block , diluted the DAB) I am still having background
>>problems. Has any one used the method where you take the sample through
>>alcohols into solvent, then back again and leave in PBS overnight before
>>you start staining if you have please reply and A. tell me if this might
>>help the background problems B.give me advice and /or methods which
>>will.The last two times I have posted a similar request no one answered so
>>someone some where this time please please answer!!!!!!        Thank you in
>>Jill McVee
>>St. Andrews Uni
>N Kenneison

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