Celltiter glo luminescent atp assay kit

Manufactured by Promega

Sourced in United States

The CellTiter-Glo luminescent ATP assay kit is a reagent-based solution that quantifies the amount of ATP present in a sample, which is an indicator of metabolically active cells. The kit uses a luciferase reaction to generate a luminescent signal proportional to the amount of ATP present, allowing for the assessment of cell viability and proliferation.

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Lab products found in correlation

Spetramax m5, by Molecular Devices (1 mentions) Spark 10m, by Tecan (1 mentions) Cytochrome oxidase activity colorimetric assay kit, by Abcam (1 mentions) 312e microplate reader, by Agilent Technologies (1 mentions)

5 protocols using celltiter glo luminescent atp assay kit

Mitochondrial Respiratory Complex Activity

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ETC activity, including the activity of NADH-ubiquinone oxidoreductase (complex I; cat. no. ab109721), succinate dehydrogenase (complex II; cat. no. ab109908), coenzyme Q-cytochrome c oxidoreductase (complex III; cat. no. ab109905) and cytochrome c oxidase (complex IV; cat. no. ab109911), were analyzed using commercial kits (Abcam) according to the manufacturer's protocol. To measure the complex I, II, III or IV activity, the detergent was added to cells to extract transmembrane proteins. Absorbance was measured on a Spetramax M5 microplate reader (Molecular Devices, LLC, Sunnyvale, CA, USA). ATP generation was detected using a CellTiter-Glo luminescent ATP assay kit (Promega Corp., Madison, WI, USA) according to the manufacturer's protocol. The supernatant was collected and added to the substrate solution. Luminescence was measured using a luminometer (Thermo Fisher Scientific Inc.).

Hao J., Du H., Liu F., Lu J.C., Yang X.C, & Cui W. (2019). Apurinic/apyrimidinic endonuclease/redox factor 1 (APE1) alleviates myocardial hypoxia-reoxygenation injury by inhibiting oxidative stress and ameliorating mitochondrial dysfunction. Experimental and Therapeutic Medicine, 17(3), 2143-2151.

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Mitochondrial ATP Quantification in Astrocytes

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Mitochondrial ATP production was quantified using the CellTiter-Glo luminescent ATP assay kit (Promega, Madison WI) according to manufacturer’s guidelines. Astrocytes were plated on 96-well black-walled assay plates and seeded at a density of 10,000 cells per well. After 24-hours of growth phase, cells received hypoxic treatment with or without reoxygenation and were allowed to equilibrate at room temperature for 30-minutes before the ATP assay was implemented. Luciferase luminescence was measured with a plate reader (BioTek). ATP was calculated in ng per 10,000 cells using an ATP standard curve. The standard curve was created immediately prior to running the assay to prevent ATP degradation. Standard curve was prepared by serial tenfold dilutions of ATP in DMEM.

Quintana D.D., Garcia J.A., Sarkar S.N., Jun S., Engler-Chiurazzi E.B., Russell A.E., Cavendish J.Z, & Simpkins J.W. (2018). Hypoxia-reoxygenation of primary astrocytes results in a redistribution of mitochondrial size and mitophagy. Mitochondrion, 47, 244-255.

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Cytochrome c Oxidase and ATP Activity Assays

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Cytochrome c oxidase activity was determined using Cytochrome Oxidase Activity Colorimetric Assay Kit (BioVision, Milpitas, CA) according to the manufacturer’s instructions. Absorbance was measured at 550 nm every 30 sec for 300 sec. Cellular ATP concentrations were measured using the CellTiter-Glo luminescent ATP assay kit, based on the luciferase/luciferin reaction, from Promega (Madison, WI, USA) according to the manufacturer’s instructions. Luminescence was assessed using a multimode microplate reader (Spark 10M, Tecan Group LTD, Mannedorf, Switzerland).

Xu L., Sun X., Griffiths B., Voloboueva L., Valdes A., Dobrenski M., Min J.J, & Stary C.M. (2023). Sexual Dimorphism in Brain Sirtuin-1 and m6A Methylated Sirtuin-1 mRNA, and in Protection with Post-Injury Anti-miR-200c treatment, after Experimental Stroke in Aged Mice. Aging and Disease, 14(3), 892-903.

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Necroptosis Induction and Measurement

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Human TNFα (hTNFα) (50 ng/mL) solution was used to induce necroptosis in FADD-deficient Jurkat cells. HT-29 and CCRF-CEM cells were pretreated with z-VAD-fmk (20 µM) and smac164 (200 nM) for 30 min before adding hTNFα (20 ng/mL) to induce necroptosis. Compounds were added to cells at the indicated concentrations and incubated for 1 day. The Cell Titer-Glo Luminescent ATP Assay kit (Promega, Madison, WI) was used to measure cell viability. Absorbance or luminescence was determined using a BioTek 312e microplate reader.

He X., Li M., Ye Z., You X., Wang J., Xiao X., Zhu G., Wei J, & Zha Y. (2023). Identification of Piperlongumine as Potent Inhibitor of Necroptosis. Drug Design, Development and Therapy, 17, 1387-1394.

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Quantifying Cellular ATP Levels

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The cellular ATP levels were quantified in 100 μL ATP assay buffer or from 20-30 mg wound skin harvested in 200 μL assay buffer and lysed with the bead shocker following the manufacturer's instructions (CellTiter-Glo luminescent ATP assay kit, Promega, Madison). The protein content from identical treated cells or tissues were determined by BCA Protein Assay kit for normalization (Young, Huang et al., 2014) .

Young G., Lin J., Cheng Y., Ho C., Kuok Q., Hsu R., Liao W., Chen C., & Chen H. (2020). Modulation of adenine phosphoribosyltransferase-mediated salvage to promote diabetic wound healing.

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