Serine/Threonine Phosphatase Assay System

• 产品编号：V2460      品牌：promega       原厂货号：V2460
• 产品分类：蛋白质检测与分析 > 酶检测和活性分析 > 磷酸酶检测
• 应用分类：

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描述：

Non-Radioactive Phosphatase Assay Systems ( 非放射性磷酸酶检测系统) 是测定蛋白磷酸酶活性的一个方便、灵活的方法。该系统确定的是游离磷酸的量，是通过反应中形成的钼酸盐: 孔雀绿:磷酸复合物的吸光度值检测出来的。该系统可与多种缓冲液组分和多种底物一起使用，包括天然的磷酸化蛋白和合成的磷酸肽。丝氨酸/苏氨酸磷酸酶分析试剂盒包含化学合成的磷酸肽RRA(pT)VA ，它是一种与多种丝氨酸/ 苏氨酸磷酸酶( 包括蛋白磷酸酶2A、2B 和2C)适合的底物。然而提供的磷酸肽不是蛋白磷酸酶1 的最适底物，因为蛋白磷酸酶1 对于底物的结构要求十分严格。

        酪氨酸磷酸酶分析试剂盒包括两种化学合成的磷酸肽, END (pY) INASL 和DADE(pY) LIPQQG，是大多数酪氨酸磷酸酶的底物。使用该系统检测磷酸酶时，有效的磷酸盐测定范围是100-4,000pmol 。该磷酸酶检测系统可以用于检测以下样品中的磷酸酶活性：细胞粗提物、组织提取物、部分纯化的样品、纯化后的样品。对于粗提物的应用而言，高浓度的磷酸需要在检测之前去除掉，使用提供的离心柱可以快速去除样品中游离的磷酸盐和其它的小分子量抑制剂。此外，独特的钼酸盐染料添加剂与钼酸盐染料溶液结合使用，可帮助暴露在酸性的钼酸盐染料溶液的蛋白质溶解，单独使用钼酸盐染料溶液时，会引起蛋白沉淀。

原厂资料：

The Non-Radioactive Phosphatase Assay Systems provide a fast, convenient and flexible alternative for measuring protein phosphatase activity. These systems determine the amount of free phosphate generated in a reaction by measuring the absorbance of a molybdate:malachite green:phosphate complex. These systems allow the use of a variety of buffer conditions and substrates, including naturally phosphorylated proteins or synthetic phosphopeptides. The Serine/Threonine Phosphatase Assay System contains the chemically synthesized phosphopetide, RRA(pT)VA, a peptide substrate that is compatible with several serine/threonine phosphatases such as the Protein Phosphatases 2A, 2B, and 2C. However the supplied phosphopeptide is a poor substrate for Protein Phosphatase 1 because of its more stringent structural requirements.

The Tyrosine Phosphatase Assay System contains two chemically synthesized phosphopeptides, END(pY)INASL and DADE(pY)LIPQQG, that serve as substrates for many protein tyrosine phosphatases. The effective range for the detection of phosphate released during an assay using the Phosphatase Assay Systems is 100–4,000pmol of phosphate. In addition to measuring phosphatase activity in partially fractionated and purified samples, the Phosphatase Assay Systems can also measure phosphatase activity in crude cell or tissue extracts. For this application, the high concentration of phosphate in these preparations is eliminated prior to performing the assay using the supplied Spin Columns, which rapidly and effectively remove free phosphate and other low-molecular-weight inhibitors from the sample. In addition, a unique Molybdate Dye Additive that is combined with the Molybdate Dye Solution aids in the solubilization of proteins exposed to the acid conditions of the Molybdate Dye Solution, which alone could potentially cause precipitation of the proteins.