[Histonet] ROOM TEMPERATURE RE: C4d antibody

From:"yichao wu"

Dear Gareth,

Thank you for your attention. Yes I agree the monoclonal antibody A213 may 
be not appliable on paraffin-embedded sections without pretreatment. I 
immerse the sections in 88% formic acid/ddH2O for 20 minutes just at room 
temperature. I think the extreme acid environment would help to uncover the 
antigen. And I also tried other pretreatments such as digestion by 0.1% 
typsin, heat pretreatment in citric acid and so on.But the signal did not 
shown up.

However, I should mention that the fixation solution is also very important 
besides the paraffin embedding. I wonder what kind of fixation solution you 
use. Actually ours is a little different from normal 10% formalin even if I 
suppose that sections fixed by 10% buffered formalin may be tried with 
formic acid as well.

Besides, I use FITC-conjugated Rabbit-anti-mouse IgG as the secondary 
antibody later in my experiment.And the fluorescence signals seem more 
easier to be observed by the eye.

As a postgraduate student myself, I am not very experienced in the field of 
immunohistology. However, I have get warm help from histonet. And  I do wish 
to communicate with more histologists here.



Yichao WU, MD
Department of Medicine
Nanjing University School of Medicine
Nanjing, China
Email: yichaowu@hotmail.com
TEL: +86-25-80860805

>From: "Gareth Bruce" 
>Subject: C4d antibody
>Date: Tue, 25 May 2004 16:25:57 +0100
>Dear Yichao WU,
>I noticed on histonet that you have experience with Quidel's antibody A213 
>to C4d.
>We have been experiencing some difficulty using this antibody in IHC with 
>paraffin sections.
>You recommend 20 minutes of 88% formic acid pretreatment.
>May I ask you what temperature this was done at?
>Thank you so very much.
>Best regards
>Gareth Bruce

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