Univ. Bordeaux, CNRS, IBGC, UMR 5095, 33000 Bordeaux, France.
STAR Protoc. 2022 May 16;3(2):101403. doi: 10.1016/j.xpro.2022.101403. eCollection 2022 Jun 17.
Lactate is a central metabolite in energy metabolism and is also involved in cell signaling and epigenetic regulations. Here, we describe an NADH-independent enzymatic assay allowing rapid, selective, and sensitive quantification of L-lactate down to the pmol range. We detail lactate extraction from intracellular and extracellular fractions, followed by total protein amount determination and enzymatic assay. This approach allows quantification of intracellular and extracellular L-lactate levels, validated by treating adherent and non-adherent cells with inhibitors of lactate transporters (MCT).
乳酸是能量代谢中的一种核心代谢物，同时也参与细胞信号转导和表观遗传调控。在这里，我们描述了一种 NADH 非依赖性的酶促分析方法，可快速、选择性和灵敏地检测低至 pmol 范围的 L-乳酸。我们详细介绍了从细胞内和细胞外部分提取乳酸，然后进行总蛋白量测定和酶促分析的方法。通过用乳酸转运蛋白（MCT）抑制剂处理贴壁和非贴壁细胞，验证了这种方法可以定量检测细胞内和细胞外的 L-乳酸水平。
