[Histonet] antibody elution solution

jincan jincan <@t> itsa.ucsf.edu
Fri Feb 20 23:04:47 CST 2004

Hi Linda,
Any solution of low pH will work to some extend. The principle is to elute 
first primary antibody (and thus also the secondary antibodies attached to 
it) from sections. Many commercial kits use 0.1N HCL. However, I found 0.1M 
glycine buffer, pH2.0 worked better (5-10 minutes each, two times). The 
buffered solution is milder to the tissue (in preserving morphology) and 
will not reveal as many new epitopes or background as 0.1N HCL. 
Nevertheless, removal of first antibody is hardly complete but only to 
reduce it to a level that would allow differentiation.
Good luck!
James Guo
Lab Manager
ImmunoVision Technologies, Co.
100 North Hill Drive, #32
Brisbane, CA 94005
Tel: 650-302-4622
Fax: 650-558-9621
Website: www.immunovisiontech.com

At 10:21 AM 2/20/2004 -0800, you wrote:
>Does anyone know if there is a commercially available reagent to elute an 
>antibody complex for a doublestaining procedure?  I am trying to double 
>label two mouse primarys using a MOM kit.  I know that Dako includes a 
>doublestain block in a kit but it is not available by itself.  I also 
>called zymed  and inquired about their doublestain enhancer. The tech rep 
>said that this product does not remove the complex and could not tell me 
>how it works....just that it might help me.  Has anyone used this product 
>before?  Thank you....Linda
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