Re: Trues Blue
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From: | "Karen D. Larison" <LARISONK@UONEURO.uoregon.edu> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
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Content-Type: | text/plain; charset="us-ascii" |
Peter S,
True Blue has been used extensively by neurochemists to label retrogradely
filled HRP-containing neurons. In these experiments, ammonium molybdate is
used to stabilize the signal. One commonly cited reference for this technique
is Olucha, J Neuroscience Methods 13, 131-138 (1985). It works quite well.
Admittedly, I only tried the method once to develop an HRP-mediated
immunohistochemical signal on frozen sections. In this experiment, the True
Blue substrate was at least 4-fold more sensitive than the DAB substrate. I
next tried the technique on whole mount embryos, and that experiment was an
abysmal failure. So I haven't pursued it further. My results comparing True
Blue and DAB were very preliminary. For instance, I hadn't titered my
antibody
out far enough to get a true picture of True Blue's sensitivity. Nor had I
tried it with a variety of antibodies. However, I've often wondered why it
isn't used more extensively in the histochemical community. I wondered about
the long-term stability of the signal. After three months, the signal is
still
robust. Does anyone know why this substrate isn't used more extensively?
Karen D. Larison
Date: Wed, 30 Dec 1998 08:21:19 +0000
From: Peter =?iso-8859-1?Q?Siesj=F6?= <peter.siesjo@neurokir.lu.se>
Subject: Trues Blue
To: histonet@pathology.swmed.edu
Hello out there,
Has anyone successfully stained something with True Blue. In our seek for a
sensitive stain for cytokine detection in frozen sections we tried it. We
were not able to get a stain for more than a couple of seconds, then it
disappeared despite different tricks recommended by the manufacturer. Or
should it be renamed False Blue?
Peter S
Peter Siesjo, M.D., Ph.D.
The Queen's Medical Center
Center for the Study of Neurological Disease
1356 Lusitana Street, U.T. 8th Floor
Honolulu, HI 96813, USA
Phone: (808) 537-7924
Phone: (home) (808) 924 1653
Fax: (808) 537-7899
Email:peter.siesjo@neurokir.lu.se
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