L-Lactate Assay Kit (Colorimetric/Fluorometric)

• Biochemical assay

PubMed

Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

Zhao et al used Lactate assay kit ab65330 and Glucose uptake assay kit 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 (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

Lab

Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

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

• FuncS

Lab

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).

• FuncS

Lab

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.

• FuncS

PubMed

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.

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/

• FuncS

Lab

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.

• FuncS

Lab

Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

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

• FuncS

Lab

Functional Studies - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

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

• Biochemical assay

PubMed

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

PubMed

Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

Shao et al used Lactate assay kit ab65330 and 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 (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

PubMed

Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

Khilazheva et al used Lactate assay kit ab65330 and ATPase assay kit 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 15,000x 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 #ab234055), using a plastic disposable pestle. After sonication, the sample was centrifugated at 10,000x 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 1000x 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.

• Biochemical assay

PubMed

Biochemical assay - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

Carreno-Florez et al used Lactate assay kit ab65330 and 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, 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.

• Other

Supplier Data

Other - L-Lactate Assay Kit (Colorimetric/Fluorometric) (AB65330)

Representative image of L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330

Components shown from left to right :

- OxiRed Probe

- L(+)-Lactate Standard

- Lactate Enzyme Mix

- Assay Buffer 2

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.