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

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 asso...

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Autores principales: Schmiedeknecht, Kira, Kaufmann, Andreas, Bauer, Stefan, Venegas Solis, Francisco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9307176/
https://www.ncbi.nlm.nih.gov/pubmed/35867690
http://dx.doi.org/10.1371/journal.pone.0271818
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author Schmiedeknecht, Kira
Kaufmann, Andreas
Bauer, Stefan
Venegas Solis, Francisco
author_facet Schmiedeknecht, Kira
Kaufmann, Andreas
Bauer, Stefan
Venegas Solis, Francisco
author_sort Schmiedeknecht, Kira
collection PubMed
description 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.
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spelling pubmed-93071762022-07-23 L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay Schmiedeknecht, Kira Kaufmann, Andreas Bauer, Stefan Venegas Solis, Francisco PLoS One Research Article 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. Public Library of Science 2022-07-22 /pmc/articles/PMC9307176/ /pubmed/35867690 http://dx.doi.org/10.1371/journal.pone.0271818 Text en © 2022 Schmiedeknecht et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Schmiedeknecht, Kira
Kaufmann, Andreas
Bauer, Stefan
Venegas Solis, Francisco
L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay
title L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay
title_full L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay
title_fullStr L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay
title_full_unstemmed L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay
title_short L-lactate as an indicator for cellular metabolic status: An easy and cost-effective colorimetric L-lactate assay
title_sort l-lactate as an indicator for cellular metabolic status: an easy and cost-effective colorimetric l-lactate assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9307176/
https://www.ncbi.nlm.nih.gov/pubmed/35867690
http://dx.doi.org/10.1371/journal.pone.0271818
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