NBT/BCIP in situ detection problem

From:Paul Klosen <klosen@neurochem.u-strasbg.fr>


Recently we have run into a nasty problem with our NBT/BCIP in situ 
detections. The in situ part seems to work fine and the alk phos detections 
is sensitive.

However, under the microscope, the precipitate is completely granular and 
seems to be located above the tissue, which is absolutely unsuitable for 
photomicrography. We have tried varying several parameters (pH, salt and 
MgCl2 conc., ..) and different lots and sources of both NBT and BCIP, but 
nothing seems to bring us back to our previous results, where we had a nice 
cell-filling precipitate, albeit a bit diffuse as expected with NBT/BCIP.

Has anyone encountered a similar problem, and how did you get rid of it ?


                                                                        (o -) O
Paul Klosen, PhD
CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
Universite Louis Pasteur  12, rue de l'Universite
F-67000 Strasbourg, FRANCE
Tel.  Fax.

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