alkaline phosphatase substrate solution

Jill Johnston johnstoj at
Thu Mar 8 15:36:41 EST 2001

Let's clear this up.  The alkaline phosphatase reaction uses BCIP
(bromo-chloro-indolyl-phosphate) and NBT (nitro-blue tetrazolium).  The BCIP is
soluble only in diethanolamine or DMSO if you have the toluidine salt.  If
however you are using the disodium salt then it is soluble in aqueous solutions
(PBS, TBS etc). NBT is soluble in DMSO.
The staining solution (50mL) is 0.2M Tris-HCl pH8.8, 0.8mM MgCl2, 5mg NBT and
2.5mg BCIP.
There are probably other solutions out there, but this is the one I use.

Jillian Johnston
Dept. of Biochemistry and Molecular Biology
University of Calgary
Phone (403) 220-3559
Fax   (403) 270-0737

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