I was wondering on average, how long does it take to stain about 15 brain slices using antibodies that detect ChAT, with a secondary antibody that is detected by AB complex (I think that's the name). The brain slices I'm using are not on 'slides', but are free floating in a staining well.

As it stands now, the protocol I'm using takes a very long while and I was wondering if the protocol I'm using is just inefficient, or if it's just average.

I'm going to admit that I'm new at IHC, and I have only tried staining a few times so plesae excuse me if I do not use the proper terms to describe my problem!

It's nice to know that there are scientists out there that actually like to chat about science! =)!

Hi and welcome to scientist solutions,
IHC should not take alot of time if the sections are already cut.
Sometimes depending upon the tissue it could take some time to prepare it I.e hard tissue as bone should be decalcified.
Usually for soft tissue it is around 4 hours give and take depending on the number of washes between stages and if the primery Ab is of good detection quality.
Usually primary and secondery ab incubation 1hr each washes 3 x 10.
If you need any more advice post and I will try to help you
We can schedule a live chat maybe it would be easier to help you

Thanks Nils!
What I'm doing now is that every time I add a reagent, and wait for the appropriate amount of time, I have to take a plastic pipette and manually drain each well while keeping the brain tissue intact.
Because of that step, I can't add reagent to all 15 wells and then drain each well, because the draining would take so long that by the time I would get to the 15th well, a the tissues would have been sitting in the reagent for too long. As a result I need to 'stagger' each row of wells when I'm adding reagent and draining reagent.

I was thinking then, that the procedure could be speeded up, if I used somekind of 'meshed' well, which would be sitting in another well. I could then add the reagents to all 15 wells, wait the appropriate time, and then pick up the wells, and let the reagent drain out, while keeping the tissue in the well.

Has anyone ever heard of a 'meshed' well plating used for IHC? I know they exsist because I've used them for another procedure that was not antibody based. So I was wondering if this is something that anyone has used?