Description
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 detection kit that is amenable to high-throughput formats and compatible with many sample types. To simplify your sample processing, the lactate detection assay uses processing methods that do not require sample centrifugation or spin columns. Simply dilute your samples, add inactivation and neutralization solutions if needed, and assay all samples directly in 96- or 384-well plates.
Luminescent Signal is Proportional to Lactate Present in Sample
Lactate Secretion by A549 Cells
Advantages of Lactate-Glo™ Assay
Quickly Detect Lactate in a Variety of Sample Types: Measure in medium, cells, tissue and plasma. The lactate detection assay requires minimal preparation steps with no need for centrifugation and spin columns.
Conduct easy in-well processing for intracellular detection.
Detect Small Changes in Lactate Levels Across a Wide Range of Concentrations: Sensitive lactate assay with broad linearity better discriminates small changes in lactate compared to colorimetric and fluorometric lactate assays.
Gain More Information Per Sample: The lactate 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.
Requires Only a Luminometer: The same plate reader that measures cell viability can be used to monitor glycolysis. No specialized instruments, plates or artificial medium are required.
Measure Multiple Metabolites from One Sample
Easily measure changes in cellular glycolysis and glutaminolysis by sampling medium over time. Direct measurement of four metabolites important to the energetic state of the cell—glucose, lactate, glutamate and glutamine—can be performed in parallel using the bioluminescent Glucose-Glo™, Lactate-Glo™, Glutamine/Glutamate-Glo™ and Glutamate-Glo™ Assays. No specialized instrument, plates or artificial medium is required. Sample processing compatible with all of the bioluminescent metabolite assays means the same sample can be used for detecting all four metabolites. This includes sample types such as culture medium, serum, plasma and tissue. The data below were produced by measuring metabolites using medium samples from the same cell cultures as described in Technical Manual TM494.
