Institute for Photonics and Advanced Sensing (IPAS), School of Biological Sciences, The University of Adelaide, Adelaide, South Australia, Australia.
Institute for Photonics and Advanced Sensing (IPAS), School of Biological Sciences, The University of Adelaide, Adelaide, South Australia, Australia.
Anal Biochem. 2021 Jun 15;623:114170. doi: 10.1016/j.ab.2021.114170. Epub 2021 Mar 15.
Colorimetric methods are convenient for the determination of inorganic phosphate. However, the acidic conditions required can complicate measurement of ATPase through non-enzymatic ATP hydrolysis. Here we present an optimized antimony-phosphomolybdate microassay for the simple and rapid detection of ATPase activity, with micromolar sensitivity. The low acidity of the color reagent results in no interference for samples containing up to 0.5-5 mM ATP, dependent on the sample volume. The assay is compatible with common assay conditions and was similar in accuracy to an established continuous method. The simplicity of this method makes it ideal for medium to high throughput applications.
比色法是一种方便的无机磷酸盐测定方法。然而，所需的酸性条件可能会使非酶促 ATP 水解复杂化，从而影响 ATP 酶的测量。在这里，我们提出了一种优化的锑磷钼酸盐微测定法，用于简单快速地检测 ATP 酶活性，具有微摩尔灵敏度。显色试剂的低酸度对含有高达 0.5-5 mM ATP 的样品没有干扰，这取决于样品的体积。该测定法与常见的测定条件兼容，并且与已建立的连续测定法具有相似的准确性。该方法的简单性使其成为中高通量应用的理想选择。
