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| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Serum, plasma, cell culture media, etc |
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Overview
For quantitative determination of L-lactate (L-lactic acid) and evaluation of drug effects on its metabolism. The assay uses FL530/585 nm for signal readout. Compatible sample input includes Serum, plasma, cell culture media, etc. Typical stated assay timing is 60 min.
Key elements and design rationale
- Readout format: FL530/585 nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Serum, plasma, cell culture media, etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 1 µM for interpreting low-signal samples.
- Feature emphasis: Sensitive and accurate. The detection limit of 1 µM and linearity up to 50 µM L-lactate in a 96-well plate assay.
Additional feature notes highlight Convenient. The procedure involves adding a single working reagent and reading the fluorescence after 60 min. Room temperature assay; High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of enzyfluo l-lactate within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
LACTATE is generated by lactate dehydrogenase (LDH) under hypoxic or anaerobic conditions. Monitoring lactate levels is, therefore, a good indicator of the balance between tissue oxygen demand and utilization and is useful when studying cellular and animal physiology. Simple, direct, and automation-ready procedures for measuring lactate concentration are very desirable. BioAssay Systems EnzyFluo™ lactate assay kit is based on lactate dehydrogenase catalyzed oxidation of lactate, in which the formed NADH reduces a probe into a highly fluorescent product. The fluorescence intensity of this product, measured at λex/em = 530/585 nm, is proportional to the lactate concentration in the sample.
Detection method
Fluorescent (FL 530/585 nm).
Detection limit and analytical sensitivity
Reported detection limit: 1 µM.
Procedures and timing
Stated procedure or timing information: 60 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify enzyfluo l-lactate in serum, plasma, cell culture media by FL530/585 nm readout.
- Compare treatment or phenotype groups using matched serum, plasma, cell culture media handling.
- Monitor time-course or pre/post changes in serum, plasma, cell culture media across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
Does heparin, EDTA or citric acid interfere with assay?
EDTA and Heparin do not interfere. 10 mM Citric acid lowers signal of 2 mM lactate by 15%.
Can this assay be used to determine lactate in cell culture supernatants that contain phenol red?
Yes, lactate assays can be performed in culture media that contains phenol red. We recommend diluting the standard in the control medium (i.e. that does not contain lactate). The detection range is 1 µM to 50 μM. If the lactate concentration in a sample is higher than 50 μM, dilute sample in the medium and repeat assays. Multiply the results by the dilution factor. The detection limit is around 1 µM. Follow the following table for standard dilutions.
Can this kit be used to measure lactic acid concentrations in cells?
Yes. To determine intracellular lactic acid concentrations, harvest cells (0.1-2 million cells per assay) in a 1.5mL tube, centrifuge 1-2 min at 3,000 rpm on a table centrifuge. Remove culture medium, wash cells quickly with cold PBS. Immediately remove any PBS. Prepare Working Reagent. Add 120 µL WR directly to the cell pellet and 80uL WR to standards in 96-wells. Vortex cell sample tube 1 min. Transfer 100 µL supernatant to 96-well. Read FL530/585nm kinetics on a plate reader.
Notes: 1. cell number. It is prudent to run several doses of cells, e.g. 0.1, 1, 2×106 cells to determine optimal cell number to be used in subsequent assays.
Working Reagent should contain a lysis reagent. BioAssay Systems will supply this reagent upon request.
How to prepare tissue homogenates for use in the assay?
For tissue sample preparation, we recommend the following:
1. Homogenize tissue in phosphate buffered saline. This can be done using a homogenizer (e.g. the Dounce type) or a grinder. If you use a lysis buffer, the buffer should not contain ascorbic acid, SDS, sodium azide, NP-40 and Tween 20.
Centrifuge to pellet any debris. This can be done at room temperature in a table centrifuge (e.g. 5 min at 14,000 rpm). The supernatant should be clear.
3. Use the supernatant for lactate assays.
Do samples containing high levels of pyruvate interfere with the assay?
Yes, high levels of pyruvate will interfere with the assay.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Multiplexed single-cell measurements of fdg uptake and lactate release using droplet microfluidics
Sengupta, D., et al (2019). Multiplexed single-cell measurements of fdg uptake and lactate release using droplet microfluidics. Technology in Cancer Research & Treatment, 18, 1533033819841066. Assay: L-Lactate in human MDA-MB-231 breast cancer cells.
Reducing l-lactate release from hippocampal astrocytes by intracellular oxidation increases novelty induced activity in mice
Vaccari Cardoso, B., et al (2021). Reducing l-lactate release from hippocampal astrocytes by intracellular oxidation increases novelty induced activity in mice. Glia. Assay: L-Lactate in HEK293 culture media.
Biological sex influences susceptibility to Acinetobacter baumannii pneumonia in mice
Pires, S., et al (2020). Biological sex influences susceptibility to Acinetobacter baumannii pneumonia in mice. JCI Insight, 5(7). Assay: L-Lactate in mice bronchoalveolar lavage fluid.
Novel molecular tools to selectively inhibit astrocyte-to-neurone L-lactate signalling
Vaccari Cardoso, B. (2021). Novel molecular tools to selectively inhibit astrocyte-to-neurone L-lactate signalling. [Thesis] Assay: L-Lactate in HEK293 cells.
Endothelial-specific FoxO1 depletion prevents obesity-related disorders by increasing vascular metabolism and growth
Rudnicki, Martina, et al (2018). Endothelial-specific FoxO1 depletion prevents obesity-related disorders by increasing vascular metabolism and growth. eLife 7: e39780. Assay: L-Lactate in mice serum.
Fibroblast bioenergetics to classify amyotrophic lateral sclerosis patients
Konrad, Csaba, et al (2017). Fibroblast bioenergetics to classify amyotrophic lateral sclerosis patients. Molecular neurodegeneration 12.1: 76. Assay: L-Lactate in human cells.
Vaginal Lactobacillus inhibits HIV-1 replication in human tissues ex vivo
Palomino, Nahui, et al (2017). Vaginal Lactobacillus inhibits HIV-1 replication in human tissues ex vivo. Frontiers in microbiology 8: 906. Assay: L-Lactate in Lactobacillus culture.
Droplet Microfluidic Platform for the Determination of Single-Cell Lactate Release
Mongersun, Amy, et al. (2016). Droplet Microfluidic Platform for the Determination of Single-Cell Lactate Release. Analytical chemistry 88.6: 3257-3263. Assay: L-Lactate in unknown cancer cells.
Compositions and methods for biological production of lactate from C1 compounds using lactate dehydrogenase transformants
Saville, Renee M., et al (2016). Compositions and methods for biological production of lactate from C1 compounds using lactate dehydrogenase transformants. U.S. Patent Application No. 14/898,948. Assay: L-Lactate in microorganism culture.
