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Atplite 1step luminescence assay system

Manufactured by PerkinElmer
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Sourced in United States, Germany

The ATPlite 1step Luminescence Assay System is a laboratory instrument designed to detect and quantify the levels of adenosine triphosphate (ATP) in biological samples. It utilizes a luminescence-based detection method to measure ATP concentrations accurately and efficiently.

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28 protocols using atplite 1step luminescence assay system

1

Cell Viability Assessment of EGCG and EC

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Toxicity of the compounds was determined by using the ATPlite 1step Luminescence Assay System of Perkin Elmer (Rodgau, Germany). Cells were incubated with EGCG or EC at different dilutions for 24 or 48 h in triplicate and analysed by adding 7.5 µl ATPlite-substrate per well, followed by detection of luciferase units with PHERAstar (BMG LABTECH, Ortenberg, Germany). The viability data are given as % relative light units of solvent (DMEM)-treated cells.
Henss L., Auste A., Schürmann C., Schmidt C., von Rhein C., Mühlebach M.D, & Schnierle B.S. (2021). The green tea catechin epigallocatechin gallate inhibits SARS-CoV-2 infection. The Journal of General Virology, 102(4), 001574.
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2

Quantifying Cell Viability by ATP Assay

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Cell viability was further assessed by measurement of adenosine triphosphate (ATP) production by means of the PerkinElmer ATPlite 1step Luminescence Assay System according to the manufacturer's protocol. After seeding and treatment as above, ARPE-19 cells were washed twice with PBS 1×, and 100 µL of buffer solution (ATPlite) was added to each well, according to the manufacturer's protocol. After 2 minutes of incubation at room temperature (shaker, 700 rpm), luminescence was assessed using the Varioskan microplate reader. Results were reported as percentage of control.
Lazzara F., Conti F., Ferrara M., Lippera M., Coppola M., Rossi S., Drago F., Bucolo C, & Romano M.R. (2023). Safety Profile of Lutein- Versus Triamcinolone Acetonide–Based Vitreous Staining. Translational Vision Science & Technology, 12(1), 5.
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3

Dose-Dependent Cell Viability Assay

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For dose response viability assays, cells were plated in 96-well tissue culture plates (1000 cells/well) in media containing DMSO or the desired concentration or each compound. Media was changed every 3 days up to a total of 9 days, at which point the ATPlite 1-Step luminescence assay system (PerkinElmer, 6016731) was used to determine ATP-dependent luminescence as an approximation of cellular viability. For cell cycle and apoptosis analysis cells were initially seeded on 10 cm dishes in media containing DMSO or 100 nM dBRD9-A and cultured/passaged in this media for a total of 9 days. For cell cycle analysis control and treated cells were harvested at 3/6/9 days and processed for FACs analysis using the BD Pharmingen BrdU Flow kit (BD, 559619) in accordance with the manufacturer’s instructions. For apoptosis analysis cells were harvested at 3/6/9 days (using Accutase to maintain cell membrane integrity) and processed for FACs analysis using the BD Annexin V Apoptosis Detection kit (BD, 556547) in accordance with the manufacturer’s instructions. Stained cells were analysed on a BD LSRFortessa Cell Analyzer and data processed using FlowJo software.
Brien G.L., Remillard D., Shi J., Hemming M.L., Chabon J., Wynne K., Dillon E.T., Cagney G., Van Mierlo G., Baltissen M.P., Vermeulen M., Qi J., Fröhling S., Gray N.S., Bradner J.E., Vakoc C.R, & Armstrong S.A. (2018). Targeted degradation of BRD9 reverses oncogenic gene expression in synovial sarcoma. eLife, 7, e41305.
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4

ATP Luminescence Assay for Compound Screening

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Measurements were performed in samples treated for 8 minutes with tested compounds, in the presence of apyrase (1 U/mL) using ATPlite 1step Luminescence Assay System (PerkinElmer).
Kaczara P., Sitek B., Przyborowski K., Kurpinska A., Kus K., Stojak M, & Chlopicki S. (2020). Antiplatelet Effect of Carbon Monoxide Is Mediated by NAD+ and ATP Depletion. Arteriosclerosis, Thrombosis, and Vascular Biology, 40(10), 2376-2390.
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5

Cell viability assay of silvestrol

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Cell toxicity of silvestrol was determined using the ATPlite 1step Luminescence Assay System from Perkin Elmer (Rodgau, Germany). Cells were incubated with silvestrol at different dilutions for 6, 24, or 48 h. By adding 7.5 µL ATPlite-substrate per well, followed by detection of the light signal with a PHERAstar FS microplate reader (BMG LABTECH, Ortenberg, Germany). The data are given as percent viability of solvent (DMSO)-treated cells.
Henss L., Scholz T., Grünweller A, & Schnierle B.S. (2018). Silvestrol Inhibits Chikungunya Virus Replication. Viruses, 10(11), 592.
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6

Cell Viability Assay with Blasticidin and Puromycin

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5x102 HN12, U87, or Cal27 cells or 1x104 SF767 cells were seeded in 5 replicate black, clear bottom 96 well plate in 6 replicate wells in complete media. After 24 hours, complete media was removed and 100μl of 10μg/ml blasticidin and 1μg/ml puromycin in DMEM + 0.5% FBS was added to each well. Baseline luminescence was measured at day 1 with the ATPlite 1step Luminescence Assay System (PerkinElmer, #6016731) on a Tecan Spark 10M. Luminescence measurements were obtained at every other day for 9 days and plotted using GraphPad Prism.
Jameson N.M., Ma J., Benitez J., Izurieta A., Han J.Y., Mendez R., Parisian A, & Furnari F. (2019). Intron 1-Mediated Regulation of EGFR Expression In EGFR-Dependent Malignancies is Mediated by AP-1 and BET Proteins. Molecular cancer research : MCR, 17(11), 2208-2220.
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7

Quantifying Cellular ATP and Mitochondrial Synthesis

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Cellular ATP was quantified with the ATPlite 1step Luminescence Assay System (Perkin Elmer), following the manufacturers’ instructions.
The mitochondrial ATP synthesis was measured in digitonin-permeabilized cells by using the luciferin/luciferase assay, as previously described (72 (link)).
Maresca A., Del Dotto V., Capristo M., Scimonelli E., Tagliavini F., Morandi L., Tropeano C.V., Caporali L., Mohamed S., Roberti M., Scandiffio L., Zaffagnini M., Rossi J., Cappelletti M., Musiani F., Contin M., Riva R., Liguori R., Pizza F., La Morgia C., Antelmi E., Loguercio Polosa P., Mignot E., Zanna C., Plazzi G, & Carelli V. (2020). DNMT1 mutations leading to neurodegeneration paradoxically reflect on mitochondrial metabolism. Human Molecular Genetics, 29(11), 1864-1881.
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8

Intracellular ATP Measurement Protocol

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Intracellular ATP levels were measured using the ATPlite, 1 step Luminescence Assay System (Perkin Elmer), a method based on the reaction of ATP with luciferase and D-luciferin. The cells were seeded in triplicates with 10,000 cells per well and treated with the indicated compounds and vehicle control for 2 or 24 hours. Luminescence was measured with Spectra Max Gemini EM luminescence microplate reader (Molecular Devices) and normalized to background levels.
Abildgaard C., Dahl C., Basse A.L., Ma T, & Guldberg P. (2014). Bioenergetic modulation with dichloroacetate reduces the growth of melanoma cells and potentiates their response to BRAFV600E inhibition. Journal of Translational Medicine, 12, 247.
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9

Cytotoxicity Assay of HA-9104

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Cells were seeded in 96-well plates in triplicate and treated with HA-9104 at various concentrations for 72 h. Cell growth was assayed by Cell Counting Kit-8 (CCK-8) (MedChem Express) at OD450 in a microplate reader (SpectraMax iD3, Molecular Devices), or by the ATPlite 1 step Luminescence Assay System (PerkinElmer) in a microplate reader. The inhibition curve was drawn by GraphPad Prism software.
Xu T., Ma Q., Li Y., Yu Q., Pan P., Zheng Y., Li Z., Xiong X., Hou T., Yu B., Liu H, & Sun Y. (2022). A small molecule inhibitor of the UBE2F-CRL5 axis induces apoptosis and radiosensitization in lung cancer. Signal Transduction and Targeted Therapy, 7, 354.
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10

Cell Viability and Clonogenic Assays for VP-16 and KU60019

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The cell viability was assessed employing the ATPlite 1-step Luminescence Assay System (PerkinElmer), as stated earlier17 (link). In brief, cells were seeded in triplicate in 96-well plates at a density of 2 × 103 cells per well, treated for 72 h with different concentrations of VP-16 or in conjunction with KU60019, and then the manufacturer's recommendations for ATPlite cell viability assaying were followed. Plots of the findings from three separate trials were generated.
For clonogenic survival assays, the cells were seeded on 60-mm dishes coated with 0.1% gelatin (Sigma, V900863) and allowed to adhere overnight. After that, the culture medium was refreshed and supplemented with DMSO, KU60019, and VP-16 alone or the combinatorial regimen for the following days. After 7–14 days, cell culture was terminated. For colony counting, colonies were photographed after being stained with coomassie brilliant blue solution.
Shu J., Wang X., Yang X, & Zhao G. (2023). ATM inhibitor KU60019 synergistically sensitizes lung cancer cells to topoisomerase II poisons by multiple mechanisms. Scientific Reports, 13, 882.
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