L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay

PLoS One. 2022 Jul 22;17(7):e0271818. doi: 10.1371/journal.pone.0271818. eCollection 2022.


Background: In recent times, the study of metabolic pathways has become inevitable and predominant for a variety of research fields as cancer biology and immunology. L-lactate as a product of anaerobic glycolysis has shown to be an important indicator of the cellular metabolic status and can be associated with diverse cellular effects. For this reason, L-lactate assay kits are of high demand when metabolic effects need to be considered. Nevertheless, commercially available kits are not affordable if multiple samples must be evaluated.

Principal finding: In this work, we develop an easy and cost-effective colorimetric assay for quantification of L-lactate suitable for cells with low or high L-lactate production based on LDH activity and suitable for 96 well-plate format. Using different metabolic regulators, we demonstrate the capacity of the assay to detect and quantify L-lactate from the supernatant of HeLa cancer cell line. Furthermore, we validate the assay against a commercially available kit by demonstrating no significant difference between both assays. Finally, we show that the assay is capable of quantifying L-lactate in primary cells such as hPBMCs that were stimulated with toll-like receptor ligands and treated with different metabolic regulators.

Conclusion: We herein present an easy custom assay that is suitable for cells with low and high L-lactate production at very low cost compared to commercially available kits. These advantages of the custom assay can simplify the research in the field of metabolism and related fields.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Colorimetry*
  • Cost-Benefit Analysis
  • Glycolysis
  • L-Lactate Dehydrogenase / metabolism
  • Lactic Acid* / metabolism


  • Lactic Acid
  • L-Lactate Dehydrogenase

Grants and funding

This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 114933180 – TR84 - C10 to S.B. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.