I use this chromagen combination for detection of in situ PCR reaction
products and as an indicator for alk. phosphatase activity in bone
(osteoblasts). The only time I had difficulty with the color remaining
through dehydration and clearing in organic solvents was when the pH of the
buffer solution was not at 9.5 or the magnesium concentration was not
adequate. As with many chemical reactions, pH and ion concentration are
critical to the formation of the permanent colored product. My two cents,
Good luck, Donna Montague, M.S.
Physiology/Biophysics and Orthopaedic Surgery
University of Arkansas for Medical Sciences
From: James Mubiru [mailto:firstname.lastname@example.org]
Sent: Thursday, July 12, 2001 1:23 PM
I am replying to a posting I saw on histonet regarding
NBT/BCIP. I have used this chromogen a few times. At
first I was told that I could not dehydrate with
alcohol and claer with xylene if I used it. But since
I was used to dehydrating and clearing using alcohol
and xylene I went ahead and did it with NBT/BCP. I did
not not see any fading of color. The color came out
very well. I would also like to hear from others who
have used it.
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