[Histonet] Re: [IHCRG] HRP-labeled primary antibodies

[michael.ho <@t> sickkids.ca]

Personally, considering the low sensitivity of the direct method, I would
just treat your antibody as if it where not conjugated, and do a species
specific polymer or ABC/LSAB method. This way you can avoid Tyramide
amplification unless really necessary....Like any new antibody, you will
have to optimize your method to give you correct specificity and
sensitivity...........

regards

Michael  Ho, MLT
Resource Technologist
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


                                                                           
             Kim Merriam                                                   
             <kmerriam2003 <@t> yah                                             
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             2009-05-21 08:41                                      Subject 
             AM                        [IHCRG] HRP-labeled primary         
                                       antibodies                          
                                                                           
             Please respond to                                             
             kmerriam2003 <@t> yaho                                             
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Hi All,

Has anyone had experience doing IHC on FFPE tissues with HRP-labeled
primary antibodies?  I was wondering what the best way to detect them would
be.  I assume that going strait to DAB would not work, since no
amplification is there.  I was thinking of using a biotinyl tyramide step
to amplify the signal.

Also, do you think the final antibody concentration would need to be higher
than with traditional, unlabeled primaries?

Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA