Direct determination of L-Lactate produced by glycolysis in cell samples. Screening of glycolysis inhibitors.

• Fast and sensitive, convenient
• Use of 5 µl sample
• Linear detection range up to 10 mM L-lactate in 96-well plate assay
• Room temperature assay, no 37 °C heater is needed
• High-throughput, "Add-mix-read" type assay
• Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day
• Method: OD565 nm

The procedure involves adding a single working reagent, and reading the absorbance after 30 minutes. Glycolysis is one of the major metabolic pathways cells undergo to produce energy and results in the production of pyruvate. One of the eventual fates of pyruvate from this process is lactate dehydrogenase converting it to L-lactate via lactic acid fermentation allowing L-lactate to serve as an indicator of glycolysis.

Glycolysis assay kit is based on measuring the production of L-Lactate from glycolysis in cells. L-Lactate that is secreted into the cell media is quantified using a coupled reaction involving the lactate dehydrogenase catalyzed oxidation of L-lactate that generates pyruvate and NADH which reduces a formazan dye. The intensity of the reduced dye, measured at 565 nm, is directly proportional to the L-lactate concentration in the sample, which in turn is directly proportional to the glycolytic rate of the cells. Samples: Cell media.