DAB staining reagents - (Jul/06/2007 )
I am new to the area of immunohistochemistry.
I want to stain human breast tissues with DAB using
mouse monoclonal antibodies.
Could you recommend which system I should
first try? Is it better using a kit?
shy of expaining the whole concept of immunohistochemistry to you i think its better if you say what systems / kits you are considering
I am now considering ABC Staining system sold by Snta Cruz.
Have you used this kit?
By the way I am going to used frozen sectios.
I'll be honest i'm not the greatest fan of santa cruz stuff as they can be a bit unpredictable.
with frozen sections you could try the following
fix with methanol, acetone, both (1:1) or parafomaldehyde
epitope retrieval with triton x
block with serum of the secondaries host animal (goat anti rabbit - block with goat)
(its a while since i did dab - is it attached to the secondary then activated?)
anyway the point is you buy the stuff separatly thus when you next do ihc you simply add to your now growing and adaptable kit (look up vector for secondaries, nuclear stains and mounting medium - you probably use dpx now i think on it)
because frozen sections arnt fixed on the slide in the oven you might consider apes coated slides to stop the sections floating off
(my current ihc is all fluorescant so its a bit different but some things stay the same)
I'm also new in IHC and I'm using the ABC Elite system, but from Vector Laboratories. It works really well, so I'd recommend it.
we also use the ABC system from vector labs. Its pretty good.