RE: normal human serum

From:Gayle Callis

One can also buy primary antibodies conjugated to biotin (Jackson
Immunoresearch has many of these), as does Pharmingen, others.  

However, you need to have biotinylated isotype matched controls or an
irrelvant biotinylated antibody of same IgG (immunoglobulin) type as the
negative control.  

The DAKO ARK kit is designed for this purpose but I am not sure it is
designed for human work, give them a call too.   


At 11:52 AM 1/25/02 -0800, you wrote:
>     Hi Carol, there are two parts to this response the first is technical
>and the second is commercial.   About doing IHC with human antibody on
>human tissues, 
>the best approach is to use the same approach that many mouse-on-mouse kits
>This is to prelabel the antibody with biotin-tagged proteins (usually an
>anti-antibody Fab fragment).  
>The procedure is: 
>To your prediluted antibody prep add an anti-human biotinylated Fab or
>related proteins (like anti-human Fc receptors). Incubate to allow binding
>to the primary antibody. Unrelated proteins (Albumin, etc.) will not be
>labeled. Excess lab  The labeled antibody is then applied to the section.
>The primary antibody will bind to its antigen, but this primary antibody
>now has biotin stuck onto it. The bound antibody is detected by the
>addition of streptavidin-enzyme conjugate. N    
>Now the commercial stuff, 
>we have performed human on human ihc services with pretty good success and
>have created a human-on-human kit based on our experience with the
>human-on-human services as well as the mouse-on-mouse iso-IHC kit that we
>sell so if you are in  Abizar 
>   > -----Original Message----- 
>> From: Carol Chang [] 
>> Sent: Friday, January 25, 2002 10:59 AM 
>> To: 
>> Subject: normal human serum 
>> Thank you all for the response of anti-human secondary  
>> antibody request. 
>> But I got another problem I need your help. I am trying to do  
>> human Ab on  
>> human tissue IHC. So I also need normal human serum to block  
>> the secondary  
>> Ab before the complex is applied to sample. Could you tell me  
>> what's the  
>> appropriate and safe way to get normal human serum except  
>> getting it from  
>> our own body that I am scared to do? 
>> Thank you and the histonet is always the place I can rely on. 
>> Carol 
>> _________________________________________________________________ 
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Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)

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