Hey Brett,

good to hear from you -- hope all is well with you.  ya, i thought about the
tyramide thing --- the AP is doing well, but i just want to pep it up a bit
--- "several companies do offer "alkaline phosphatase enhancer" --- which
one have you had experience with and which do you prefer?

take care,
rob


Original Message-----
From: Connolly, Brett M
To: 'Robert Geske'; 'histonet@pathology.swmed.edu'
Sent: 9/28/01 9:10 AM
Subject: RE: NBT/BCIP

Rob,
Assuming you don't want to go the tyramide route, which is a
possibility,
several companies do offer "alkaline phosphatase enhancer".
Brett

Brett M. Connolly, Ph.D.
Merck Research Laboratories
Dept. of Pharmacology
WP26A-3000
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-652-2075
e-mail. brett_connolly@merck.com
 

-----Original Message-----
From: Robert Geske [mailto:RGeske@lexgen.com]
Sent: Friday, September 28, 2001 8:30 AM
To: 'histonet@pathology.swmed.edu'
Subject: NBT/BCIP


i need to enhance the signal provided by the reaction product of an
alk-phos
detection system using NBT/BCIP.  this is for  mRNA in situ
hybridization of
a dig labeled probe.  i have an excellent AP conjugated anti-dig
antibody,
but i just need to increase the signal (development temp was 37).  i
have
done a series of development times, but the signal needs to be stronger.
i
have done the development on an automated system, but the "store bought"
reagent is extremely expensive.  i want to replicate the steps after the
hybe, stringency washes, and antibody application is done one line (i
just
want to do the signal development off line). i know the hybe is good and
the
mRNA copy number is detectable by non-isotopic methods.  is anyone using
something comparable to nickel or copper in a NBT/BCIP chromagen mix to
boost the signal?

thanks in advance,
rob

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