Cromogen/Substrate IHC question -Reply

From:Tony Henwood <>

Experience shows that the optimal titre for one ABC technique should be
pretty close for another. The best way is to run the antibodies on your
positive controls as well as tissues you know should be negative.
Positive staining should be similar, but you might find an increase in
background staining. In this case you will need to retitre the localising
antibody and if background is still there then you might have to revert
back to old demonstration technique, try a different demo technique or
put up with the increased background.

Just some thoughts

Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital Westmead,
Locked Bag 4001, Westmead, NSW 2145.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

>>> "Bennett, Catherine (Katie)" <> 7/February/2001
07:23am >>>
Advice needed:

If I switch an IHC procedure over from an ABC-HRP kit to an ABC-Alk
kit, I shouldn't have to adjust my primary and secondary antibody
concentrations, right?  (I want to change cromogen labels from DAB to


Catherine "Katie" Bresee Bennett
Sr. Research Technologist
Lovelace Respiratory Research Institute
Albuquerque, New Mexico

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