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L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 is a quantitative, addition-only assay with just one 30 min incubation step at room temperature. Readout on any colorimetric (570 nm) or fluorometric (Ex/Em 360nm/440nm) plate reader.

- Cited in over 250 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.


Images

Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330), expandable thumbnail
  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330), expandable thumbnail
  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330), expandable thumbnail
  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330), expandable thumbnail
  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330), expandable thumbnail

Publications

Key facts

Detection method
Colorimetric/Fluorometric
Sample types
Tissue Lysate, Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Cell Lysate
Assay type
Quantitative
Reactive species
Mammals
Range
0.001 - 10 mM
Assay time
40m
Sensitivity
> 0.001 mM

Associated Products

Select an associated product type

2 products for Alternative Product

What's included?

2000 Test
Components
Assay Buffer 2
20 x 25 mL
L(+)-Lactate Standard
20 x 100 µL
Lactate Enzyme Mix
20 x 1 Vial
OxiRed™ Probe
20 x 0.2 mL

Recommended products

L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 is a quantitative, addition-only assay with just one 30 min incubation step at room temperature. Readout on any colorimetric (570 nm) or fluorometric (Ex/Em 360nm/440nm) plate reader.

- Cited in over 250 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

Key facts

Detection method
Colorimetric/Fluorometric
Sample types
Tissue Lysate, Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Cell Lysate
Assay type
Quantitative
Reactive species
Mammals
Range
0.001 - 10 mM
Assay time
40m
Assay Platform
Microplate reader
Sensitivity
> 0.001 mM

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 provides a convenient means for detecting L(+)-Lactate in biological samples such as blood, cells, culture mediums, fermentation mediums, etc. There is no need for pretreatment or purification of samples.

How the assay works
In the lactate assay protocol, lactate specifically reacts with an enzyme mix to generate a product, which interacts with a lactate probe to produce color (570 nm) and fluorescence (at Ex/Em = 535/587 nm).

Lactate assay protocol summary

  • - Add samples (deproteinized) and standards to wells.
  • - Add reaction mix and incubate for 30 min at room temp.
  • - Analyze with microplate reader.

Related Lactate assay products
Alternative L-Lactate assay kits offer different readout modes/wavelengths and sensitivity/range:

L(+)-Lactate is the major stereo-isomer of lactate formed in human intermediary metabolism and is present in blood. D(-)-Lactate is also present but only at about 1-5% of the concentration of L(+)-Lactate.

We also provide D-Lactate assay kit D-Lactate Assay Kit (Colorimetric) ab83429.

Lactate assay methods
There are two lactate assay methods established in biological research:

  • a) The most commonly used method, where lactate dehydrogenase processes lactate in the presence of NAD+ to produce pyruvate and NADH. NADH then reduces a probe to form a colorimetric or fluorometric readout. This method, with some improvements, is used in L-Lactate Assay Kit (Colorimetric) ab65331 and in L-Lactate Assay Kit (Fluorometric, High Sensitivity) ab169557.
  • b) The alternative method, where lactate oxidase processes lactate to produce pyruvate and hydrogen peroxide. This is followed by oxidation of a substrate by a peroxidase using the hydrogen peroxide to produce a colorimetric or fluorometric readout. This method is used in ab65330.

Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K607 Lactate Colorimetric/Fluorometric Assay Kit. K607-100 is the same size as the 100 test size of ab65330.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

L-Lactate also known as lactate is a byproduct of anaerobic glycolysis where it plays an important role in energy metabolism. L-Lactate is a small molecule with a molecular mass of approximately 90.08 g/mol. It forms in various tissues like muscle cells during intense exercise when oxygen levels are low. This process leads to a conversion of pyruvate to lactate by the action of the enzyme lactate dehydrogenase (LDH) which is present in many tissues with higher expressions in muscles and heart.

Biological function summary

L-Lactate acts as a signaling molecule which affects cellular functions and contributes to metabolic regulation. It is not part of a complex but serves as an important intermediate in metabolic pathways. L-Lactate provides energy to cells by converting back to pyruvate in the presence of oxygen which then enters the citric acid cycle. This conversion and its utilization as energy play important roles in balancing cellular energy demands especially under hypoxic conditions.

Pathways

L-Lactate links to critical processes like glycolysis and the Cori cycle. During glycolysis pyruvate may convert to L-Lactate under anaerobic conditions to regenerate NAD+ necessary for glycolysis to continue. In the Cori cycle lactate produced by anaerobic glycolysis in muscles is released into the bloodstream and transported to the liver. There it converts back to glucose supporting gluconeogenesis. These processes highlight the close involvement of L-Lactate with proteins such as lactate dehydrogenase and pyruvate kinase.

Associated diseases and disorders

L-Lactate is associated with conditions like lactic acidosis and cancer. Lactic acidosis characterized by high lactate levels can occur due to oxygen deprivation or mitochondrial dysfunction. Meanwhile cancer cells often show enhanced glycolysis and lactate production known as the Warburg effect facilitating their growth. Proteins like hypoxia-inducible factor 1-alpha (HIF-1α) and lactate dehydrogenase (LDH) are key players in these conditions influencing lactate metabolism and potentially serving as therapeutic targets.

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12 product images

  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail
    Image from Lai D., PLiS One 8(9), Fig 6C. Doi: 10.1371/journal.pone.0075625. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    HaCaT cells are grown in normoxia for 3 days after infection and medium was tested to determine lactate concentration using L-Lactate assay kit (ab65330). HaCaT cells were infected with AdGFP-18E2 or AdGFP, transfected with Ctrl or HIF-1α siRNA.

  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Relative signal (RFU) in unfiltered human plasma (dilution 1:200) and mouse urine (dilution 1:200), comparing L-lactate signals with background readings ((-); no enzyme) after 10 minutes of incubation (duplicates +/- SD).

  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Lactate measured in cell culture supernatants showing quantity (μmol) per mL of tested sample. Samples were diluted 40-160 fold and measured fluorometrically.

  • Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Lactate measured in biological fluids showing quantity (μmol) per mL of tested sample. Samples were diluted 10-40 fold and measured fluorometrically.

  • Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Diagram showing the principles of the L-Lactate assay method.

  • Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Khilazheva et al used Lactate assay kit ab65330 with mouse hippocampal homogenates as part of analysis of the phenotype of an NLRP3 KO mouse.

    Determination of lactate levels in the hippocampal homogenates obtained from adult (5 months) and aged (14 months) mice of different genotypes: WT (wild-type mice (C57Bl/6)) and NLRP3 KO (knockout mice for the NLRP3 gene), nmol/µg of protein. **—p ≤ 0.01. n = 5 to 8 mice per group. Values are presented as M ± SEM. Lactate concentration in the hippocampus was analyzed using an enzymatic method followed by a colorimetric measurement.

    Lactate concentration in the hippocampus was analyzed using an enzymatic method followed by a colorimetric measurement. A commercially available kit (L-Lactate Assay Kit, ab65330, Abcam, UK) was utilized for this purpose. The sensitivity of the kit is reported to be greater than 0.001 mM. Protein concentration measurements were determined using the Bio-Rad protein assay kit with bovine serum albumin as standards (Bio-Rad). The resulting protein concentrations were expressed in μg/μL. To calculate the concentration of lactate in each sample, the obtained values in nmol/μL were divided by the corresponding protein concentration. Consequently, the lactate concentration in the homogenate was presented as nmol/μg of protein.

  • Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Carreno-Florez et al used Lactate assay kit ab65330 and Glucose assay kit Glucose Assay Kit ab65333 to investigate glycolytic flux and the Warburg effect in cultured human ΔF508/ΔF508 cystic fibrosis airway epithelial cells (CFBE41o-, hereafter called CF AECs).

    Glucose consumption in the basolateral medium of IFN-β-stimulated or influenza virus and respiratory syncytial virus (RSV)-infected CF AECs measured by colorimetric assay. L-lactate concentration measured in apical secretions from CF AECs during IFN-β stimulation or RSV infection measured by colorimetric assay. For all experiments n ≥ 3. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Glucose consumption (Abcam, Glucose Assay Kit ab65333) and lactate secretion (Abcam, ab65330) were measured after 18 h of IFN-β stimulation or 72 h of RSV infection using colorimetric assays. Glucose consumption was determined by subtracting the glucose concentration in basolateral medium from the glucose concentration in the feeding medium. For lactate secretion, MEM without phenol red was added apically for 18 h during IFN-β stimulation or for the last 24 h of RSV infection and was collected to determine the extracellular lactate concentration.

  • Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Zhao et al used Lactate assay kit ab65330 and Glucose uptake assay kit Glucose Uptake Assay Kit (Colorimetric) ab136955 to investigate in cell culture supernatants of mouse primary liver cells and UBR7 over-expressing BEL-7402 cells.

    WT and Alb-Cre;UBR7fl/fl mouse primary liver cells were analysed for glucose uptake and lactate secretion levels. Glucose uptake and lactate secretion levels were detected in UBR7 overexpressing BEL-7402 cells. Data are shown as mean +/- SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.

    Lactate assay kit (ab65330) and Glucose Uptake Assay (Glucose Uptake Assay Kit (Colorimetric) ab136955) were purchased from Abcam. The detection of lactate, glucose uptake and ATP levels were carried out according to the method recommended by the kit.

  • Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Shao et al used Lactate assay kit ab65330 and Glucose assay kit Glucose Assay Kit ab65333 to investigate the impact of Salvigenin, a potential therapeutic) on glycolysis with cell culture supernatant of HepG2 cells.

    Salvigenin repressed HCC cell aerobic glycolysis. Salvigenin (25 µM, 50 µM, 100 µM) was utilized to treat HepG2 cells. The level of glucose uptake. The level of lactate production.

    The supernatant of the mediums of HepG2 cells was harvested. The floating cells and cell debris were removed through 5 min of centrifugation (1000 g, 4 °C). Then, the supernatant was harvested. As instructed by the manufacturer, the glucose uptake detection kit (Glucose Assay Kit ab65333, Abcam, USA) and the lactate production detection kit (ab65330, Abcam, USA) were taken to determine the levels of glucose uptake and lactate generation.

  • Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330), expandable thumbnail

    Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    Khilazheva et al used Lactate assay kit ab65330 and ATPase assay kit ATPase Assay Kit (Colorimetric) ab234055 to investigate how an inducible Slc4a11 KO mouse leads to corneal edema by disruption of the pump and barrier functions of the corneal endothelium (CE).

    Changes in the pump function in the inducible Slc4a11 KO. Stomal lactate content; relative values versus control; mean ± SEM, n = 3, *: p < 0.05. Na+-K+ ATPase activity at 14 days of Tm treatment; mean ± SEM, n = 3 (Ctrl) and n = 5 (Tm), *: p < 0.05.

    Stromal Lactate

    Corneas were obtained, and the epithelium and endothelium were removed. Individual stromas were placed in pre-weighed Eppendorf tubes, pulverized in liquid nitrogen, and homogenized in 30 µL of PBS using a plastic disposable pestle. The sample was centrifuged at 15,000× g for 15 min at 4 °C. The supernatant was recovered. The remaining pellet was dried at 60 °C in a vacuum centrifuge for two hours, then weighed (dry weight). Lactate was measured in the supernatant, n 3 for Ctrl and Tm, using a fluorescent kit (Abcam #ab65330, Cambridge, UK) according to the manufacturer’s instructions.

    Na+-K+ ATPase Activity

    Two corneal endothelial-Descemet membrane (CEDM) peelings from the same mouse were pooled and homogenized in 30 µL assay buffer, provided in the ATPase Assay kit (Abcam #ATPase Assay Kit (Colorimetric) ab234055), using a plastic disposable pestle. After sonication, the sample was centrifugated at 10,000× g for 10 min at 4 °C. The supernatant was recovered, and the phosphates in the sample were depleted by incubation in 40 µL of PiBind resin (Innova Biosciences #501-0015, Montluçon, France) for 15 min at room temperature in a rotary device. After centrifugation at 1000× g for two minutes, the sample was recovered, and ATPase activity was measured in 5 µL of sample in the presence or absence of 1 mM ouabain. Na+-K+ ATPase activity (n 3 for Ctrl and Tm) was obtained by subtracting the activity in presence of ouabain from the total activity. Protein was measured using the BCA method.

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