L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric/Fluorometric
- Platform: Microplate reader
- Assay time: 40 min
- Sample type: Cell culture supernatant, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Lysate, Urine
- Sensitivity: 0.001 mM
Overview
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Product name
L-Lactate Assay Kit (Colorimetric/Fluorometric)
See all L-Lactate kits -
Detection method
Colorimetric/Fluorometric -
Sample type
Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Cell Lysate, Tissue Lysate -
Assay type
Quantitative -
Sensitivity
> 0.001 mM -
Range
0.001 mM - 10 mM -
Assay time
0h 40m -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
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.
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 -
Notes
L(+)-Lactate is the major stereo-isomer of lactate formed in human intermediary metabolism and is present in blood. D(-)-Lactate is also present (see D-Lactate assay kits) but only at about 1-5% of the concentration of L(+)-Lactate.
Alternative L-Lactate assay kits offer different readout modes/wavelengths and sensitivity/range:
- this kit L-Lactate assay ab65330: colorimetric 570 nm, fluorometric Ex/Em 535/587 nm, range 0.001 mM - 10 mM
- our most popular L-Lactate assay ab65331: colorimetric 450nm, range 0.02 mM - 10 mM
- L-Lactate assay ab169557: fluorometric Ex/Em 535/587 nm, range 0.2 µM - 50 µM.Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
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. -
Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests L(+)-Lactate Standard (100 nmol/µl) Yellow 1 x 100µl Lactate Assay Buffer WM 1 x 25ml Lactate Enzyme Mix (lyophilized) Green 1 vial Lactate Probe (in DMSO, anhydrous) Red 1 x 200µl -
Research areas
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Relevance
Lactate (CH3CH(OH)COO-) plays important roles in many biological processes. Abnormal high concentration of lactate has been related to disease states such as diabetes and lactate acidosis, etc. L(+)-Lactate is the major stereoisomer of lactate formed in human intermediary metabolism and is present in blood. The lactate to pyruvate ratio reflects the redox state of the cell and describes the balance beween NAD+ and NADH, which is dependent on the interconversion of lactate and pyruvate via lactate dehydrogenase (LDH).
Images
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Functional studies - ab65330 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/
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.
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Functional Assay: 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).
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Functional Studies - L-Lactate Assay Kit (ab65330)Standard curve: mean of duplicates (+/- SD) with background reads subtracted
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Functional Studies - L-Lactate Assay Kit (ab65330)Standard curve: mean of duplicates (+/- SD) with background reads subtracted
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Functional Studies - L-Lactate Assay Kit (ab65330)Lactate measured in cell culture supernatants showing quantity (µmol) per mL of tested sample. Samples were diluted 40-160 fold and measured fluorometrically.
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Functional Studies - L-Lactate Assay Kit (ab65330)Lactate measured in biological fluids showing quantity (µmol) per mL of tested sample. Samples were diluted 10-40 fold and measured fluorometrically.
