Hi Nasir,
I use Biocare reagents (DAB, Warp Red and Ferengi Blue) for my multicolour
stuff.
Even though the Vector blue fluoresces you should still see what you need to
see in brightfield. If you're worried about losing your signal through
solvents you could probably just airdry before coverslipping, that's what I do
(using Ecomount). The xyelene is only a problem after you put the chromogen
on, not before.
Looks like you have 2 rabbit and 1 rat Ab for your Abs, it may be possible to
do a triple stain with that with a rat and rabbit in the first round and the
other rabbit in the 2nd or you should be able to combine at least a rat and
rabbit in a double simultaneous or 2 rabbits in a double sequential stain
depending on your Abs and how robust they are. If you don't have access to
spectral imaging though it may be safer going with doubles so you can pull
everything apart by eye.
Is that CD8 Ab behaving ok for you? Looks like it has a ton of background in
the images. If you're having issues with it you may be able to combine your
CD4 Ab with the CD3 SP7 clone to show the presumptive CD8s.
I haven't worked with any of those Abs but I do do multicolour IHC in both
human and mouse tissues so if you need help sorting out protocols let me know,
of course I can only speak to Biocare reagents for this as that's what I
usually use.
Katy Milne
Deeley Research Centre
Message: 2
Date: Fri, 4 Nov 2016 15:36:54 +0000 (UTC)
From: Nasir Abbas <shinwar...@yahoo.com>
To: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] Suggestion for Dual staining Kit/reagent
Message-ID: <735822620.281524.1478273814...@mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8
Hi All
Can some one suggest dual staining reagents for IHC-P (mouse liver tissue) for
my primary antibodies: ab183685, GTX53126 and ab25377.I was suggested vectors
labs products (following table) but I am not clear about Vector Blue reagent
SK-5300 that whether it can be used in IHC-P, because it features shows it is
fluorescent. Moreover it is sensitive to xylene and Histoclear may be used as a
clearing reagent in this case, which makes me even more worried. Any suggestion
and tips for my dual staining?Table:
| Pri-Ab | Vector Lab Products- Suggested |
| ab183685 | MP-7401-15 |
| GTX53126 | SK-4105 |
| ab25377 | MP-5404-15 |
| ? | SK-5300 |
?
Sincerely?Nasir Abbas
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
