Name: L-Lactate Assay Kit (Colorimetric)
SKU: ab65331
Rating: 5 (2 reviews)

L-Lactate Assay Kit (Colorimetric) ab65331 is a quantitive, addition-only assay with one 30-min room temperature incubation

- Cited in over 300 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) (AB65331), expandable thumbnail

Publications

Key facts

Detection method: Colorimetric
Sample types: Tissue Lysate, Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Cell Lysate
Assay type: Quantitative
Range: 0.02 - 20 mM
Assay time: 30m
Sensitivity: = 0.02 mM

Associated Products

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Target data

See full target information L-lactate

What's included?

100 Test

Components

Assay Buffer 12

1 x 25 mL

Developer Solution III

1 x 1 Vial

Enzyme Mix XV

1 x 1 Vial

L(+)-Lactate Standard

1 x 100 µL

Key facts

Detection method: Colorimetric
Sample types: Tissue Lysate, Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Cell Lysate
Assay type: Quantitative
Range: 0.02 - 20 mM
Assay time: 30m
Assay Platform: Microplate reader
Sensitivity: = 0.02 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) (ab65331) uses an assay protocol where lactate is oxidized by lactate dehydrogenase to generate a product which interacts with a probe to produce a color (λmax = 450 nm).

The kit detects L(+)-Lactate in biological samples such as serum or plasma, cells, tissues, cell culture and fermentation media.

Lactate assay protocol summary

- Add samples 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 assay L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330: colorimetric 570 nm, fluorometric Ex/Em 535/587 nm, range 0.001 mM - 10 mM
- This kit, our most popular L-Lactate assay kit ab65331: colorimetric 450nm, range 0.02 mM - 10 mM
- L-Lactate assay kit L-Lactate Assay Kit (Fluorometric, High Sensitivity) ab169557: fluorometric Ex/Em 535/587 nm, range 0.2 µM - 50 µM.

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.

How other researchers are using L-Lactate Assay Kit ab65331
This Lactate assay kit has been used in publications in a variety of sample types, including:

- Human: THP-1 cell lysates¹, MDA-MB-231 and HepG2 cell culture lysates², cell culture supernatant (HepG2, A549, Huh7, PC3, LN229, HeLa)³, brain tissue⁴
- Mouse: brown adipose tissue lysate⁵, thymic lymphoma tissue⁶, cell culture supernatant⁷, T cell primary cell culture supernatants⁸, serum⁹, serum and muscle¹⁰
- Bovine: cumulus cell culture supernatant¹¹

^{References: 1 - Tran UT and Kitami T 2019; 2 - Cui J et al 2019; 3 - Rodriguez ML et al 2018, Chen Y et al 2018, Zhang D et al 2018, Caino MC et al 2017, Birkenmeier K et al 2015; 4 - Sullivan RC et al 2019; 5 - Jeong JH et al 2018; 6 - Vara-Ciruelos D et al 2019; 7 - Fiorenzano et al 2016; 8 - Menk AV et al 2018; 9 - Deng W et al 2019, Guglielmetti C et al 2017, Kang R et al 2016; 10 - Kim HY et al 2016; 11 - Sinha et al 2017.}

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 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 L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330.

Other notes
This product was previously called K627 Biovision Lactate Colorimetric Assay Kit II. Abcam acquired BioVision in 2021.

REACH authorisation
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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9 product images

Functional Studies - L-Lactate Assay Kit (Colorimetric) (ab65331)
Linearity of dilution: concentration of L-Lactate in differently diluted (X-axis) biological samples, demonstrating a linearity of 89%-111% (concentrations corrected for by factor of dilution; duplicates; +/- SD).
Image from Kelly CE et al., PLoS Biol 11(4), fig2d. oi: 10.1371/journal.pbio.1001538. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Functional Studies - L-Lactate Assay Kit (Colorimetric) (ab65331)
Plasma lactate concentrations were determined using L-Lactate assay kit (ab65331) in Ark2C^+/+ and Ark2C^−/−(Arkadia-like gene) mice.
Functional Studies - L-Lactate Assay Kit (Colorimetric) (ab65331)
Relative signal (RFU) in unfiltered human plasma (dilution 1:8), comparing L-lactate signals with background reading (no enzyme) after 10 minutes of incubation (duplicates +/- SD).
Functional Studies - L-Lactate Assay Kit (Colorimetric) (ab65331)
Standard curve with background signal subtracted (duplicates; +/- SD).
Functional Studies - L-Lactate Assay Kit (Colorimetric) (ab65331)
Lactate Standard Curve. The assay is performed following the kit (ab65331) protocol.
Biochemical assay - L-Lactate Assay Kit (Colorimetric) (ab65331)
Diagram showing the principles of the L-Lactate assay method.
Enzyme activity assay - L-Lactate Assay Kit (Colorimetric) (ab65331)
Pyruvate Kinase Assay Kit Pyruvate Kinase Assay Kit ab83432 used with cell culture lysates, and Lactate Assay Kit ab65331 used with cell culture medium.
Li et al. used Pyruvate Kinase Assay Kit Pyruvate Kinase Assay Kit ab83432 and Lactate Assay Kit ab65331 with cultured human primary airway smooth muscle cells (ASMCs) derived from healthy donors (#CC-2576, N = 4, passage 2) and Chronic obstructive pulmonary disease (COPD) patients.
PKM2 activity is essential for metabolic reprogramming towards aerobic glycolysis in COPD-derived ASMCs. (A) Pyruvate kinase activity was measured in normal ASMCs with si-RNA mediated HHIP and/or PKM2 knockdown with normalization to protein concentration. (C) Lactate levels in the culture medium were measured in COPD-derived ASMCs transfected with si-Ctrl or si-PKM2.
Pyruvate Kinase Assay Kit (#Pyruvate Kinase Assay Kit ab83432) was purchased from Abcam.
Measurement of secreted lactate levels in cell culture media: Cells were seeded into a 96-well plate at the density of 3 × 103 cells/well. The culture medium was then collected 24 h after the seeding of cells. The lactate levels in the medium were measured by the Lactate Colorimetric Assay Kit II (#K627-100, BioVision, Milpitas, CA) [Biovision is an Abcam company, K627 is now sold as Abcam ab65331], and lactate levels were then normalized to DNA content measured in each well at the time of collection.
Biochemical assay - L-Lactate Assay Kit (Colorimetric) (ab65331)
Riedel et al used Lactate assay kit ab65331 with tissue culture media of mouse cells and mouse skin tissue extracts and showed that tumor-derived lactic acid drains to lymph nodes where it modulates the function of lymph node stromal cells.
Tumor-derived lactate is responsible for the observed changes in FRC activation and mitochondria. L-lactate concentration in CCM, B16.F10 TCM, and 4T1 tissue culture media, as measured by an enzymatic assay. n = 3 experiments with 1 to 5 replicates. L-lactate quantification in tissues collected from either Balb/c sham-treated mice (MFP) and orthotopic 4T1 tumors (4T1 tumor), or C57BL/6 sham-treated (skin) and B16.F10 melanoma tumors (B16 tumor). Measured by an enzymatic assay. n = 4 to 6 independent experiments. Representative of three independent experiments. Data are means with SEM. Significance (*, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001) was determined by unpaired two-tailed t test or one-way ANOVA with Tukey post hoc (B). B16, B16.F10; max., maximum; min, minute; LA, lactic acid.
L-Lactate was measured in conditioned medium and tissue samples using the L-Lactate Assay kit (abcam, #ab65331) according to the manufacturer's instructions. All samples were deproteinized with the Deproteinizing Sample Preparation Kit–trichloroacetic acid (TCIA; abcam, #Deproteinizing Sample Preparation Kit - TCA ab204708) before assaying.
Tissue samples were weighed into Precellys tubes prefilled with ceramic beads. The exact 6x volume (1 mg tissue/6 µL buffer) of Lactate Assay Buffer was added on ice and samples were lysed using a Precellys 24 homogeniser (Bertin Instruments).
Biochemical assay - L-Lactate Assay Kit (Colorimetric) (ab65331)
Gruszczyk et al used Lactate assay kit ab65331 to investigate the impact of anoxia in mouse primary cardiomyocyte cultures.
Cardiomyocytes were incubated for different time periods of anoxia or 120 min normoxia at 37 °C with various concentration of AR-C141990. Control buffer with 0 µM MCT1 inhibitor is shown in black, MCT1 inhibitor-containing buffer in red. The supernatant and cell pellets were then isolated and analyzed. Cell lactate concentration (pmol/105 cells) on the left y-axis and lactate fold changes to control on the right y-axis (n = 3–5 biological replicates, *P < 0.03, **P < 0.01, ***P < 0.0005, one-way ANOVA with multiple comparisons and Tukey's post hoc test.
Lactate was measured in cell supernatants or in cell pellets. For analysis of the intracellular lactate content, the cardiomyocytes were washed and lysed in 200 µL assay buffer/105 cells by scraping them down and repeating three freeze/thaw cycles and vigorous vortexing in between these cycles. The samples were centrifuged for 2 min at 14,000 g at 4 °C and the supernatant kept for deproteinization. Therefore, 25 µL 5 M perchloric acid were added to each tube, followed by 47 µL of 3 M potassium hydroxide to neutralize the samples. After centrifugation for 5 min at 14,000 g at 4 °C, the supernatant was used to determine lactate concentrations using the colorimetric Abcam Lactate Assay Kit (ab65331). According to the manual instructions, a standard curve was prepared via serial dilution of the kit's standard concentrate in assay buffer. 50 µL of each standard, blank or sample were pipetted into a CoStar 96- well plate. Then, a reaction master mix was prepared containing 46 µL assay buffer, 2 µL substrate mix and 2 µL enzyme mix per reaction. 50 µL of this master mix were added to each well and the reaction was incubated for 30 min at 400 rpm at room temperature. The output was measured on a microplate reader (CLARIOstar PLUS from BMG Labtech) at OD 450 nm.