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Elanecdotario - Phosphatase coupled sulfotransferase assay: r&d systems. This phosphatase coupled assay is universal to all sulfotransferases that use paps as the donor substrate the assay is quantitative and allows accurate kinetic analysis it is a convenient and cost effective assay because no radioisotope and no separation of products and substrates are required. Universal sulfotransferase activity kit. Universal sulfotransferase activity kit catalog number: ea003 the protocol presented is for a 96 well format materials provided are sufficient for two 96 well microplates or equivalent. Golgi resident pap specific 3' phosphatase coupled. A universal phosphatase coupled sulfotransferase assay is described here in this method, golgi resident pap specific 3' phosphatase gpapp is used to couple to a sulfotransferase reaction by releasing the 3' phosphate from pap the released phosphate is then detected using malachite green reagents the enzyme kinetics of gpapp have been determined, which allows calculation of the coupling. Golgi resident pap specific 3´ phosphatase coupled. Scheme of phosphatase coupled sulfotransferase assay during the sulfotransferase reaction, the donor substrate, paps, is converted to pap in the coupling reaction, the 3´ phosphate on pap is removed by gpapp the free inorganic phosphate can subsequently be detected by malachite green reagents. Recombinant human heparan sulfate 2 o 6xoirwudqvihudvh 6 67. Prepare reaction mixture containing 0 4 mm paps, 6 mg ml heparan sulfate, and 20 ?g ml coupling phosphatase 3 in assay buffer 4 dilute rhhs2st1 to 20 µg ml in assay buffer. Zhengliang wu google scholar citations. Golgi resident pap specific 3´ phosphatase coupled sulfotransferase assays b prather, cm ethen, m machacek, zl wu analytical biochemistry 423 1 , 86 92 , 2012. Adenosine 3´ phosphate 5´ phosphosulfate lithium salt. Adenosine 3´ phosphate 5´ phosphosulfate lithium salt has also been used in a study to assess the golgi resident pap specific 3´ phosphatase coupled sulfotransferase assays other notes. Carbohydrate sulfotransferase 10 chst10 springerlink. Abstract the sulfotransferase with the official gene name carbohydrate sulfoctransferase 10 chst10 is involved in the biosynthesis of the hnk 1 carbohydrate epitope, originally described as a differentiation antigen specifically expressed on human natural killer hnk cells abo and balch 1981. Recent advances in sulfotransferase enzyme activity assays. One novel st coupled assay system involves the well studied enzyme, aryl sulfotransferase iv ast iv , which transfers a sulfo group from paps to aryl alcohols in this assay, paps is continuously regenerated in st catalyzed reactions by using p nitrophenyl sulfate pnps as a sulfo group donor, facilitating colorimetric monitoring of the pnp product at 400 nm scheme 4. Pdf a versatile polyacrylamide gel electrophoresis based. Scheme of electrophoresis based sulfotransferase assay the assay mainly consists of four steps first, spike sulfotransferase reaction with pap 35 s.
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