Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Nishi-Ku, Fukuoka, 819-0395, Japan.
Graduate School of Systems Life Sciences, Kyushu University, 744 Motooka, Nishi-Ku, Fukuoka, 819-0395, Japan.
Anal Sci. 2024 Nov;40(11):2075-2080. doi: 10.1007/s44211-024-00631-6. Epub 2024 Jul 20.
Effect of drugs on the intracellular activity of lactate dehydrogenase (LDH) has been measured by using water-soluble tetrazolium (WST). Because the assay is usually conducted in the presence of dead cells, net activity of live cells is not evaluated. Here, we reported the assay of the net intracellular LDH activity of live cells by counting the live cells using fluorescent staining of nucleus. By using a deep red fluorescent dye, dual measurements of fluorescence signal of nucleus and absorbance of WST could be conducted with transparent 96-well-plates. We found that conventional assay in the presence of dead cells overestimate the effect of drugs on the LDH activity.
采用水溶性四唑盐（WST）测定了药物对乳酸脱氢酶（LDH）细胞内活性的影响。由于该测定通常在死细胞存在的情况下进行，因此无法评估活细胞的净活性。在这里，我们通过使用核荧光染色来计数活细胞，报道了活细胞内 LDH 活性的净测定方法。通过使用深红色荧光染料，可以用透明的 96 孔板进行细胞核荧光信号和 WST 吸光度的双重测量。我们发现，在存在死细胞的常规测定中，药物对 LDH 活性的影响被高估了。
