From:Robert Geske

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,

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