I am attempting a dual staining technique with one HRP labelled antibody
and one AP labelled antibody. For technical reasons the AP antibody
needs to be incubated last. Could somebody please suggest as to whether
I should develop the HRP substrate (in this case DAB) and then incubate
with the AP antibody or if I should perform all the antibody incubations
and then develop the substrates.
I am staining cryofixed sections.
If somebody could also suggest a good nuclear counter stain it would be
appreciated. My HRP substrate is DAB and I am using "Fast Blue BB Base"
as my AP substrate.
Thanks in advance for your trouble.
David Smith (dasmith at yoyo.cc.monash.edu.au)
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David Smith
Honours Student
Monash University Biochemistry Department
