||Enolase (EC 18.104.22.168), also called 2-Phospho-D-glycerate hydrolase, is a key enzyme in glycolysis. It converts 2-phosphoglycerate into phosphoenolpyruvate (PEP). It also catalyzes the reverse reaction, PEP to 2-phosphoglycerate under anabolic conditions during gluconeogenesis. In mammals, three genes encode for the Enolase α, β and γ isoforms. These isoforms combine together to form both homo and heterodimeric complexes resulting in five isozymes, three of which are commonly found in human tissues. Enolase α, also called enolase 1, exists in variety of tissues which can undergo glycolysis. Besides its role in glycolysis, increased Enolase α activity is associated with tumor invasion and metastasis; therefore, screening for novel and specific inhibitor of enolase α inhibitor may be of great interest for tumor treatment. In Human Enolase α Inhibitor Screening Kit, Enolase α catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate, which is subsequently used to generate an intermediate product. The intermediate product stoichiometrically reacts with OxiRed; probe generating a colorimetric signal (OD 570 nm). In the presence of Enolase α inhibitor, the reaction is impeded. An enolase α Inhibitor Control is included to compare the efficacy of the sample inhibitors. The assay is simple, fast, high-throughput adaptable and can be completed in less than 1 hr.