[Histonet] RE: Over lapped flouresence in double staining
Dear Dr. Sohail,
You didn't write us what detection system was used for this double
staining. The fact you observed overlap of SMA and vWF (which isn't
very likely indeed) makes your detection system a bit suspect. To my
opinion the most straightforward (and safe) ways to doublestain these
A: cocktail of SMA (mouse) + vWF (rabbit), followed by a secondary
cocktail composed of goat anti-mouse/FLUO-1 + goat anti-rabbit/FLUO-2
B: cocktail of SMA, clone 1A4 (mouse IgG2a) + vWF , clone F8/86 (mouse
IgG1), followed by a secondary cocktail composed of goat anti-mouse
IgG2a/FLUO-1 + goat anti-mouse IgG1/FLUO-2
Lots of success!
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
NL-1105 AZ Amsterdam
Date: Tue, 18 Apr 2006 19:00:05 -0700 (PDT)
From: sohail ejaz
Subject: [Histonet] Over lapped flouresence in double staining
I am doing double staining of endothelial cells and smooth muscles of
the blood vessels with vWF and SMA.
While doing double staining I have encountered overlapping of both
antibodies and I cant see a clear differentiation between the
fluorescence of these two antibodies.
Could you please guide me how to over come this problem.
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