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Inorganic and organic phosphate measurements in the nanomolar range.

Analytical biochemistry (1987-02-15)
P P Van Veldhoven, G P Mannaerts
ABSTRACT

A procedure, based on the complex formation of malachite green with phosphomolybdate under acidic conditions, to measure inorganic orthophosphate in the nanomolar range is described. The addition of polyvinyl alcohol is required to stabilize the dye-phosphomolybdate complex. The advantages of the assay are simplicity, stability of the reagents, and high sensitivity. Due to the high permissible acidity in the assay (0.9 N H2SO4), the method can be adapted easily to measure nanomolar amounts of phosphate, liberated from organic compounds like phosphoproteins and phospholipids after wet digestion.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Alkaline/Acid Phosphatase Assay Kit (R-R-A-pS-V-A), Alkaline/Acid Phosphatase Assay Kit is routinely used to detect phosphatase activity by either dephosphorylation of the phosphopeptide (RRApSVA) or hydrolysis of pNPP.
Sigma-Aldrich
PTP Assay Kit 1, PTP Assay Kit 1 can be used to detect PTP-1B activity by dephosphorylation of the phosphopeptide (RRLIEDAEpYAARG) using malachite green detection, or by hydrolysis of pNPP.
Sigma-Aldrich
Ser/Thr Phosphatase Assay Kit 1 (K-R-pT-I-R-R), Ser/Thr Phosphatase Assay Kit 1used to detect PP2A activity by either dephosphorylation of the phosphopeptide.