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Adenosine 5 triphosphate bioluminescent somatic cell assay kit

Manufactured by Merck Group
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The Adenosine 5'-triphosphate bioluminescent somatic cell assay kit is a laboratory tool used to measure the levels of adenosine 5'-triphosphate (ATP) in somatic cells. It utilizes bioluminescent technology to quantify the ATP present in a sample.

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Lab products found in correlation

N luminometer, by Promega (2 mentions) White 96 well plate, by Corning (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions)

Close spelling variants of this product

Adenosine 5′-triphosphate (ATP) bioluminescent somatic cell assay kit Bioluminescent Somatic Cell Assay Kit Adenosine 5′-triphosphate Bioluminescent assay kit Adenosine

4 protocols using adenosine 5 triphosphate bioluminescent somatic cell assay kit

1

Cellular ATP Depletion Effects on Breast Cell Phenotypes

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Normal, tumorigenic, and metastatic breast cells were incubated for 3h in either phenol red-free MEGM (Lonza) as the control condition, or with low azide (2mM sodium azide and 2mM deoxy-D-glucose) or high azide (8mM sodium azide and 50mM 2-deoxy-D-glucose) added to this MEGM medium. Cells were trypsinized, resuspended in the same media, and assayed in a white 96-well plate (Corning, Corning, NY) using the Adenosine 5′-triphosphate bioluminescent somatic cell assay kit (Sigma-Aldrich cat. no. FLASC). Cell samples were read using a well integration time of 10 s in a Perkin Elmer Enspire 2300 luminometer within 10min of luciferase/luciferin assay mix addition. The percentage of ATP remaining in low- and high-azide-treated samples was determined from a ratio of their averaged relative luminescence units (RLU) compared to those of control samples. Each cell type and treatment combination was assayed in technical triplicate, and for normal and tumorigenic cells, biological duplicate as well.
After high-azide treatment of the cells on poly-D-lysine-coated flasks, normal cells appeared to have ruffled lamellipodia, and normal, tumorigenic, and metastatic cells did not round up normally upon trypsinization, but peeled off the flask surface retaining their spread morphology.
Smelser A.M., Macosko J.C., O’Dell A.P., Smyre S., Bonin K, & Holzwarth G. (2015). Mechanical properties of normal versus cancerous breast cells. Biomechanics and modeling in mechanobiology, 14(6), 1335-1347.
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2

Measuring ATP Levels in Cells

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ATP levels after CDN or control agent treatment were determined by Adenosine 5’-triphosphate Bioluminescent Somatic Cell Assay Kit (Sigma-Aldrich, FLASC). Briefly, 106 cells were seeded in 6-well plate 24 h prior to the treatment. Other than the control group, cells were incubated with CDN, SPN, TPA or ATN224 at concentrations indicated in the figure legends for 24 h in complete medium or medium without FBS supplement. For ROS scavenger experiments, various concentrations of MitoTEMPO was added to the medium for co-incubation. Cells were then trypsinized and resuspended in ultrapure water for assay. The luminescence signals were measured by Turner Biosystems 20/20n luminometer. The ATP concentration of each group was determined based on the ATP standard assay (n=3 per condition ATP production measurement for each cell line).
Cui L., Gouw A.M., LaGory E.L., Guo S., Attarwala N., Tang Y., Qi J., Chen Y.S., Gao Z., Casey K.M., Bazhin A.A., Chen M., Hu L., Xie J., Fang M., Zhang C., Zhu Q., Wang Z., Giaccia A.J., Gambhir S.S., Zhu W., Felsher D.W., Pegram M.D., Goun E.A., Le A, & Rao J. (2020). ED SUM: Triple-negative breast cancer is inhibited by depleting mitochondrial copper in mice.: Mitochondrial copper depletion suppresses triple-negative breast cancer in mice. Nature biotechnology, 39(3), 357-367.
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3

Antibiotic Cytotoxicity Evaluation in Kidney Cells

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Human Kidney 2 cells (ATCC CRL-2190) were obtained from the American Type Culture Collection and maintained in high-glucose Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 1% of penicillin-streptomycin combo solution and 10% fetal bovine serum (Sigma-Aldrich, Poole, UK). Cells were cultured at 37 °C in 5% CO2 saturated air, replacing the culture media every 48 hours. Cells were passaged upon reaching 80% confluency, and seeded in 96-well flat bottom plates at a density of 1.2 × 104 cells/well in 200 μl of DMEM medium without serum. After a further 24 hours’ culturing, cells were exposed to the antibiotic under test for 6 hours and cytotoxicity assessed by measuring lactate dehydrogenase leakage with the CytoTox-ONETM Homogenous Membrane Integrity Assay (Promega, USA), and by monitoring intracellular ATP levels using the Adenosine 5′-triphosphate bioluminescent somatic cell assay kit (Sigma-Aldrich, UK).
Nass N.M., Farooque S., Hind C., Wand M.E., Randall C.P., Sutton J.M., Seipke R.F., Rayner C.M, & O’Neill A.J. (2017). Revisiting unexploited antibiotics in search of new antibacterial drug candidates: the case of γ-actinorhodin. Scientific Reports, 7, 17419.
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4

Measuring ATP Levels in Cells

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ATP levels after CDN or control agent treatment were determined by Adenosine 5’-triphosphate Bioluminescent Somatic Cell Assay Kit (Sigma-Aldrich, FLASC). Briefly, 106 cells were seeded in 6-well plate 24 h prior to the treatment. Other than the control group, cells were incubated with CDN, SPN, TPA or ATN224 at concentrations indicated in the figure legends for 24 h in complete medium or medium without FBS supplement. For ROS scavenger experiments, various concentrations of MitoTEMPO was added to the medium for co-incubation. Cells were then trypsinized and resuspended in ultrapure water for assay. The luminescence signals were measured by Turner Biosystems 20/20n luminometer. The ATP concentration of each group was determined based on the ATP standard assay (n=3 per condition ATP production measurement for each cell line).
Cui L., Gouw A.M., LaGory E.L., Guo S., Attarwala N., Tang Y., Qi J., Chen Y.S., Gao Z., Casey K.M., Bazhin A.A., Chen M., Hu L., Xie J., Fang M., Zhang C., Zhu Q., Wang Z., Giaccia A.J., Gambhir S.S., Zhu W., Felsher D.W., Pegram M.D., Goun E.A., Le A, & Rao J. (2020). ED SUM: Triple-negative breast cancer is inhibited by depleting mitochondrial copper in mice.: Mitochondrial copper depletion suppresses triple-negative breast cancer in mice. Nature biotechnology, 39(3), 357-367.
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