Bacterial alkaline phosphatase reaction

The fractions containing cross-linked species are pooled and lyophi-lized several times to remove the volatile salt. The residue is dissolved in approximately 100 1 of 10 mM ammonium carbonate at pH 7.5 and mixed with unlabeled tRNA. This mixture is then digested for 4-5 hr at 37° with Tl ribonuclease (about 5 units per Aseo of total tRNA ) and bacterial alkaline phosphatase (about 5 per A ea of total tRNA). After digestion, the reaction mixture is extracted with an equal volume of... 

Alkaline phosphatase catalyzes the dephosphorylation of a mmber of artificial substrates ( ) including 3-glycerophosphate, phenylphosphate, p-nitrophenylphosphate, thymolphthalein phosphate, and phenolphthalein phosphate. In addition, as shown recently for bacterial and human enzymes, alkaline phosphatase simultaneously catalyzes the transphosphorylation of a suitable substance which accepts the phosphoryl radical, thereby preventing the accumulation of phosphate in the reaction mediim (25). 

Immunoassay and nucleic acid probe assays Labels. recApoaequorin, firefly luciferase, marine bacterial luciferase, Vargula luciferase Detection reactions, alkaline phosphatase label (luciferin-O-phosphate/firefly luciferase), glucose 6-phosphate dehydrogenase label (marine bacterial luciferase/NADH FMN oxidoreductase reaction)... 

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