[Histonet] RE: Same Host Ab(fluorescence IHC) HELP...!!!
I have been testing this Jackon protocol once and could not figure out
a good staining either. I presume that this type of protocol only
works under "ideal" conditions. That means if you have exactly
titrated all your reagents perfectly. Just a little too much of one
reagent will result into non-specific, or better to say "unwanted"
As mentioned on Histonet, the approach to biotinylate one primary via
the Dako ARKit (or even both primaries via the Zenon kit), is much
Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
NL-1105 AZ Amsterdam
phone: +31 20 5665631
fax: +31 20 6960389
----- Original Message -----
Date Thu, 03 Feb 2005 10:59:32 -0800
Subject [Histonet] Same Host Ab(fluorescence IHC) HELP...!!!
Here is the problem that is giving me a lot of grey hairs even
I am only 26. Its making me crazy and I am in deep despair. I cant
on like this.. Please help..! :'(
I have to do Fluorescent based IHC on two proteins on old
fixed skin tissue. The first antibody telomerase is from Mouse. I
goat antimouse Fab2 fragments of Alexa488 for the secondary
The second Ab that I have to stain is melanoma cells and I use
Melanoma cocktail from Biocare Medicals for this purpose. This
again from Mouse and has several IgG subtypes like IgG1K, IgG2a,
etc. I use Goat antimouse TRITC as my secondary for this step.
! ; t
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