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Next we add subtrates, which in the presence of the enzyme will convert into a dye. For example, in the case of using alkaline phosphatase (ALP), a solution of BCIP/NBT is added (5-bromo-4-chloro-3-indolyl phosphate + nitro blue tetrazolium). The dye created under catalytic influence of the enzyme, sediments on the well bottom. After visible spots develop, colour reaction can be halted through thorough washing and drying the plate. More...
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