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Atp bioluminescence assay kit

Manufactured by Merck Group
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Sourced in United States, India

The ATP bioluminescence assay kit is a laboratory equipment product that measures the concentration of adenosine triphosphate (ATP) in a sample using bioluminescence technology. The core function of this kit is to quantify the amount of ATP present, which is an essential molecule involved in various cellular processes.

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Close spelling variants of this product

ATP assay kit (67 citations) ATP Bioluminescence Assay Kit CLS II (17 citations) ATP Bioluminescence Assay Kit HS II (10 citations) ATP-luciferase-based bioluminescence assay kit (5 citations) Bioluminescence assay kit (4 citations) Bioluminescence assay (3 citations)

34 protocols using atp bioluminescence assay kit

1

Measuring Cellular ATP and ROS Levels

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Cellular ATP and ROS levels were measured as described previously [32 (link)]. Cells cultured in a 6-well plate after various treatments, were lysed and cellular ATP levels were measured with ATP Bioluminescence Assay Kit (Sigma). For measuring glycolysis-mediated ATP production, cells were treated with 2 μM rotenone for 24 hours before measurements.
Cui Y., Qin L., Wu J., Qu X., Hou C., Sun W., Li S., Vaughan A.T., Li J.J, & Liu J. (2015). SIRT3 Enhances Glycolysis and Proliferation in SIRT3-Expressing Gastric Cancer Cells. PLoS ONE, 10(6), e0129834.
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2

Cell ATP Bioluminescence Assay

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Cells were trypsinized, and >105 cells were subjected to ATP assays using the ATP bioluminescence assay kit (Sigma-Aldrich) according to the manufacturer's instructions.
Lee B., Ahn Y., Kang S.M., Park Y., Jeon Y.J., Rho J.M, & Kim S.W. (2015). Stoichiometric expression of mtHsp40 and mtHsp70 modulates mitochondrial morphology and cristae structure via Opa1L cleavage. Molecular Biology of the Cell, 26(12), 2156-2167.
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3

Measurement of ATP and GSH in RBCs

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After the RBCs were exposed to PS-NPs for 3 h, the intracellular levels of ATP and GSH were measured. ATP levels in RBCs were analyzed using diluted RBCs reacted with an ATP assay mix, following the protocol for the ATP bioluminescence assay kit (Sigma-Aldrich). To detect GSH levels, we followed the method described by the manufacturer of the GSH-Glo™ GSH Assay (Promega). After centrifugation, we removed the supernatant and added GSH-Glo reaction buffer. We centrifuged the suspended RBCs and transferred the supernatant to a new tube. Diluted samples were reacted with GSH-Glo reagent buffer and luciferin detection reagent under dark conditions. We measured the luminescence of ATP and GSH using the EnSpire (PerkinElmer).
Kim E.H., Choi S., Kim D., Park H.J., Bian Y., Choi S.H., Chung H.Y, & Bae O.N. (2022). Amine-modified nanoplastics promote the procoagulant activation of isolated human red blood cells and thrombus formation in rats. Particle and Fibre Toxicology, 19, 60.
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4

Mitochondrial ATP Production Assay

Cited 2 times
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After the measurement of ADP-stimulated respiration of complex I and complex II, the incubation buffer containing mitochondria was taken from the respiration chamber and immediately supplemented with the ATP assay mix diluted 1:5 (ATP Bioluminescence Assay Kit, Sigma-Aldrich, St. Louis, MO, USA). Mitochondrial ATP production was determined immediately after each respiration measurement and compared with ATP standards using a 96-well white plate in a spectrofluorometer (SpectraMax® M3, Molecular Devices, San Jose, CA, USA) at 560-nm emission [23 (link),27 (link),28 (link)].
Rosa F.L., de Souza I.I., Monnerat G., Campos de Carvalho A.C, & Maciel L. (2023). Aging Triggers Mitochondrial Dysfunction in Mice. International Journal of Molecular Sciences, 24(13), 10591.
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5

Luminometric ATP Determination in Mitochondria

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Luminometric method [using ATP bioluminescence assay kit (Sigma, St. Louis, MO, United States)] was used to determine ATP levels according to the provided protocol. Mitochondrial samples were assayed for ATP content using the ATP dependence of the light emitting luciferase-catalyzed oxidation of luciferin. ATP concentration was calculated according to a standard curve and related to protein content (Wang et al., 2014 (link)).
Husain I., Akhtar M., Madaan T., Vohora D., Abdin M.Z., Islamuddin M, & Najmi A.K. (2018). Tannins Enriched Fraction of Emblica officinalis Fruits Alleviates High-Salt and Cholesterol Diet-Induced Cognitive Impairment in Rats via Nrf2–ARE Pathway. Frontiers in Pharmacology, 9, 23.
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6

Intracellular ATP Quantification

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Cells were cultured in six-well plates. After various treatments, cells were lysed by 0.5% Triton X-100 in 100 mM glycine buffer, pH 7.4. Intracellular ATP levels were assayed with an ATP bio-luminescence assay kit (Sigma) based on the luciferase-catalyzed oxidation of d-luciferin.
Zhao L., Liu Z., Jia H., Feng Z., Liu J, & Li X. (2015). Lipoamide Acts as an Indirect Antioxidant by Simultaneously Stimulating Mitochondrial Biogenesis and Phase II Antioxidant Enzyme Systems in ARPE-19 Cells. PLoS ONE, 10(6), e0128502.
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7

Oxidative Stress Biomarkers Assay

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T-AOC and the activities of SOD1 and SOD2 were analyzed using Jiancheng Biochemical detection kits according to the corresponding kit protocols. ATP content was detected using an ATP bioluminescence assay kit (Sigma) as previously described.48 (link)
Gao J., Feng Z., Wang X., Zeng M., Liu J., Han S., Xu J., Chen L., Cao K., Long J., Li Z., Shen W, & Liu J. (2017). SIRT3/SOD2 maintains osteoblast differentiation and bone formation by regulating mitochondrial stress. Cell Death and Differentiation, 25(2), 229-240.
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8

Quantifying Extracellular ATP Release in mRECs

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Cells were prepared for ATP release assays. The ectoATPase inhibitor β, γ-methylene-ATP (300 lM; Sigma-Aldrich Corp) was added 15 min prior to each stimulation. Extracellular ATP in mRECs culture medium was measured 15 min and 45 min after LPS stimulation using an ATP bioluminescence assay kit (Sigma-Aldrich Corp). Luminescence was quantifified using a luminometer (TD 20/20; Turner Designs, San Jose, CA, USA). Concentrations of ATP were calculated using an ATP standard curve.
Evaluation of LPS in peripheral blood supernatant was a quantitative test for Gram-negative bacterial endotoxin (Lonza, USA; Cat No. QCL-1000).
Kong H., Zhao H., Chen T., Song Y, & Cui Y. (2022). Targeted P2X7/NLRP3 signaling pathway against inflammation, apoptosis, and pyroptosis of retinal endothelial cells in diabetic retinopathy. Cell Death & Disease, 13(4), 336.
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9

Quantification of Total Cellular ATP

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To measure total ATP content, 5x105 H9C2 cells were suspended and lysed in a buffer containing 0.22 M sucrose, 0.12 M mannitol, 40 mM tricine, pH 7.5 and 1 mM EDTA at 4˚C for 10 min. The total lysate was analyzed using ATP Bioluminescence Assay kit (Sigma-Aldrich; Merck KGaA) according to the manufacturer's instructions and quantitatively measured using the Optocomp I BG-1 luminometer (GEM Biomedical, Inc.).
Tang Q., Xiong W., Ke X., Zhang J., Xia Y, & Liu D. (2020). Mitochondria-associated protein LRPPRC exerts cardioprotective effects against doxorubicin-induced toxicity, potentially via inhibition of ROS accumulation. Experimental and Therapeutic Medicine, 20(4), 3837-3845.
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10

Evaluation of Rosuvastatin and Piracetam Interaction

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Rosuvastatin (RSV) and piracetam (PCT) were provided as a gift sample by Sun
Pharmaceutical Industries Limited and Arbro Pharmaceuticals Limited (India),
respectively. JC-1 (5, 5, 6, 6- tetrachloro-1,1,3,3- tetraethyl benzimidazolyl
carbocyanine iodide) and ATP bioluminescence assay kit were procured from Sigma
Aldrich (India). BCA protein assay kit was purchased from Span Diagnostics
Limited, Gujarat, India. All other reagents used were of analytical grade.
Double-distilled water was used throughout the experimental work.
Husain I., Akhtar M., Madaan T., Abdin M.Z., Islamuddin M, & Najmi A.K. (2018). Rosuvastatin alleviates high-salt and cholesterol diet-induced cognitive impairment in rats via Nrf2–ARE pathway. Redox Report : Communications in Free Radical Research, 23(1), 168-179.
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