Lactate-Glo™ Assay

How the Lactate Assay Works

The Lactate-Glo™ Assay couples lactate oxidation and NADH production with a bioluminescent NADH detection system. Lactate dehydrogenase uses lactate and NAD+ to produce pyruvate and NADH. In the presence of NADH a pro-luciferin Reductase Substrate is converted by Reductase to luciferin, which is then used in a luciferase reaction to produce light. The Lactate-Glo™ Assay contains an L-lactate-selective lactate dehydrogenase to confer specificity for L-lactate, the major stereoisomer found in mammalian cells.

When Lactate Detection Reagent, which contains lactate dehydrogenase (LDH), NAD+, Reductase, Reductase Substrate and Luciferase, is added to a sample containing lactate at a 1:1 ratio, the enzyme-coupled reactions start and run simultaneously. The luminescent signal is proportional to the amount of lactate in the sample and increases until all lactate is consumed at which time a stable luminescent signal is achieved.

Meets Your Throughput and Workflow Requirements

The Lactate-Glo™ Assay is a versatile system that is amenable to high-throughput formats and compatible with many sample types. Samples may require upfront sample processing, including dilutions, to fit into the linear range of the assay. They may also require inactivation of endogenous enzyme activity/deproteinization and NAD(P)H degradation. To simplify sample processing, methods for rapid enzyme inactivation and NAD(P)H degradation are provided that are compatible with 96- and 384-well plate formats and do not require sample centrifugation or spin columns.

Advantages for Lactate-Glo™ Assay

Quickly Detect Lactate in a Variety of Sample Types: Measure in medium, cells, tissue and plasma. The lactose detection assay requires minimal preparation steps with no need for centrifugation and spin columns.

Conduct easy in-well processing for intracellular detection.

Measure Lactate Across a Wide Range of Concentrations: Broad linearity up to 3 logs with simple sample measurement when using the lactate assay.

Detect Small Changes in Lactate Levels: Wide assay window with a S/Bmax > 200 better discriminates small changes in lactate compared to colorimetric and fluorometric lactate assays.

Gain More Information Per Sample: The lactose detection assay along with the other bioluminescent metabolite detection assays are amenable to measuring multiple metabolites from the same sample. Multiplex with cell viability assays for normalization.