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Atp detection assay kit luminescence

Manufactured by Cayman Chemical
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The ATP Detection Assay Kit-Luminescence is a laboratory tool used to detect the presence and quantify the amount of adenosine triphosphate (ATP) in a sample. It utilizes a luminescence-based detection method to provide a sensitive and reliable measurement of ATP levels.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Tiron, by Thermo Fisher Scientific (1 mentions) Ros glo, by Promega (1 mentions) Antimycin a, by Merck Group (1 mentions) Centro xs3 lb 960, by Berthold Technologies (1 mentions) Cm h2dcfda, by Merck Group (1 mentions) Jc 1 staining, by Cayman Chemical (1 mentions) Sense microplate reader, by Hidex (1 mentions) Glutamate assay kit fluorometric, by Abcam (1 mentions) Synergy h1 plate reader, by Agilent Technologies (1 mentions) Victor x4 multilabel plate reader, by PerkinElmer (1 mentions) Triton x 100, by PerkinElmer (1 mentions) Glutaraldehyde, by Electron Microscopy Sciences (1 mentions) Easysep mouse na ve cd8 t cell isolation kit, by STEMCELL (1 mentions) Mojosort mouse anti apc nanobeads, by BioLegend (1 mentions) Siinfekl peptide, by Biosynth (1 mentions) Recombinant murine il 15, by Thermo Fisher Scientific (1 mentions) Poly d lysine, by Merck Group (1 mentions) Seahorse xf cell mito stress test kit, by Agilent Technologies (1 mentions) Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions)

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ATP Detection Assay Kit (18 citations) ATP assay kit (6 citations) ATP detection kit (2 citations)

11 protocols using atp detection assay kit luminescence

1

Quantifying ATP in Knee Synovial Fluid

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Synovial fluid of the knee joint was absorbed by a strip of paper filter (2 mm x 10 mm). Volume of synovial fluid was determined by the weight increase of the paper strip. Materials absorbed to the strips were eluted with 20 μl of distilled water. Amounts of ATP in the eluates were determined using a luminescence ATP detection assay kit (Cayman Chemical Company, Ann Arbor, MI). ATP concentration was calculated from the amount of ATP and the volume of synovial fluid.
Habuchi H., Izumi M., Dan J., Ushida T., Ikeuchi M., Takeuchi K, & Habuchi O. (2021). Bone marrow derived mast cells injected into the osteoarthritic knee joints of mice induced by sodium monoiodoacetate enhanced spontaneous pain through activation of PAR2 and action of extracellular ATP. PLoS ONE, 16(6), e0252590.
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2

Measuring ROS and ATP in HT22 Cells

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For ROS measurement, HT22 cells were treated with carboxy-H2-dichlorodihydrofluorescein diacetate (DCFDA) reagent (C400, Thermo Fisher Scientific, Waltham, MA) in Hank’s balanced salt (HBSS) medium (14175-095, Gibco, Thermo Fisher Scientific, Waltham, MA) for 30 min, washed and treated with 100 µM of ROS inducer antimycin A (J63522, Sigma-Aldrich, St. Louis, MO) alone or with different concentrations of AC102 or DMTU (D188700, Sigma-Aldrich, St. Louis, MO) or Tiron (A0447328, Thermo Fisher Scientific, Waltham, MA) for 20 min in DMEM. After the incubation, DCFDA fluorescence (Ex: 485 nm; Em: 535 nm) was measured at ClarioStar cell analyzer (BMG Labtech, Ortenberg, Germany) and blanks from cell-free wells were subtracted. For cell-free ROS assays, 1 µM H2O2 was incubated with AC102 or with DMTU for 2 h and levels of H2O2 was determined by luminescence-based assay (ROS-Glo, G8820, Promega, Madison, WI) and normalized to protein levels. For ATP measurement, HT22 cells were treated with varying concentrations of AC102 for 4 h. Total ATP production was measured with the Luminescence ATP Detection Assay Kit (Cayman Chemical, 700410) according to the manufacturer’s instructions. All data were normalized against the protein levels determined by the bicinchoninic acid (BCA) assay (23223, Pierce, Thermo Fisher Scientific, Waltham, MA) under the same experimental conditions.
Rommelspacher H., Bera S., Brommer B., Ward R., Kwiatkowska M., Zygmunt T., Theden F., Üsekes B., Eren N., Nieratschker M., Arnoldner C., Plontke S.K., Hellmann-Regen J, & Schlingensiepen R. (2024). A single dose of AC102 restores hearing in a guinea pig model of noise-induced hearing loss to almost prenoise levels. Proceedings of the National Academy of Sciences of the United States of America, 121(15), e2314763121.
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3

Quantifying Cellular Metabolites

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AGS (1 × 106) cells were seeded in 12-well plates. After 24 h treatment with various formulations, the cells were incubated with A-CoA (Acetyl Coenzyme A) ELISA Kit (Elabscience, Houston, TX, USA), D-Lactic Acid/Lactate Colorimetric Assay Kit (Elabscience), and ATP Detection Assay Kit-Luminescence (Cayman), separately. The relative luminescence levels of ATP and the relative absorbance levels of acetyl-CoA and lactate were detected using a TECAN ELISA reader at luminescence or absorbance wavelength of 450 nm and 530 nm, respectively.
Lo Y.L., Wang T.Y., Chen C.J., Chang Y.H, & Lin A.M. (2022). Two-in-One Nanoparticle Formulation to Deliver a Tyrosine Kinase Inhibitor and microRNA for Targeting Metabolic Reprogramming and Mitochondrial Dysfunction in Gastric Cancer. Pharmaceutics, 14(9), 1759.
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4

ATP-release Assay for SH-10-TC Cells

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For ATP-release from SH-10-TC cells, ATP-detection assay kit - Luminescence was used according to the manufacturer’s instructions (Cayman Chemical). SH-10-TC cells (0.25 × 105 cells/200 μl/wells) (96 well plate) were cultured in the presence or absence of 5 mM 2DG or 5 μM GO-Y022. After 24h of incubation at 37°C and 5% CO2, change the supernatants to PBS and incubate another 10 min at 37°C and 5% CO2. The luminescence according to ATP-release from tumor cells in each treatment (PBS-samples) was read using CentroXS3 LB960 (Berthold Technologies, Bad Wildbad, Deutschland).
MaruYama T., Miyazaki H., Lim Y.J., Gu J., Ishikawa M., Yoshida T., Chen W., Owada Y, & Shibata H. (2023). Pyrolyzed deketene curcumin controls regulatory T cell generation and gastric cancer metabolism cooperate with 2-deoxy-d-glucose. Frontiers in Immunology, 14, 1049713.
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5

Islet Glucose-Stimulated Insulin Secretion Assay

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Isolated islets were dissociated with Trypsin-EDTA (0.25%) for 7 min at 37 °C. The digestion was completed by pipetting, then the islets were washed three times with KRBB +2.8 mM glucose solution and centrifuged at 500×g for 2 min. The pellet was resuspended in KRBB +2.8 mM glucose solution, and islets starved for 1 h at 37 °C. During the starvation period, islets were pretreated with CM-H2DCFDA (500 nM; Sigma) and 10 μM TPPU or 2 μM EETs mixture. Then islets were washed and seeded into 96 well plates, and CM-H2DCFDA fluorescence was measured before and after glucose stimulation following the manufacturer’s instructions. Additionally, the ATP concentration was evaluated using the ATP Detection Assay Kit-Luminescence (Cayman Chemical). Briefly, twenty size-matched islets were collected at each time point during ex-vivo GSIS and snap-frozen. Islets were lysed with 10 μL of ATP detection sample buffer, and ATP was measured following the manufacturer’s instructions, and data were normalized to the protein level. Moreover, the cAMP was determined using HitHunter® cAMP Assay for Small Molecules (Discover X). Twenty-five size-matched islets were collected before and after ex-vivo GSIS and snap-frozen, and the cAMP was measured following the manufacturer’s instructions.
Koike S., Hsu M.F., Bettaieb A., Chu B., Matsumoto N., Morisseau C., Havel P.J., Huising M.O., Hammock B.D, & Haj F.G. (2021). Genetic deficiency or pharmacological inhibition of soluble epoxide hydrolase ameliorates high fat diet-induced pancreatic β-cell dysfunction and loss. Free radical biology & medicine, 172, 48-57.
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6

Quantifying Cellular ATP Levels

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Absolute levels of ATP per 105 cells were determined using the ATP Detection Assay Kit—Luminescence (Cayman Chemical, Ann Arbor, MI, USA). Briefly, hematopoietic cell lineages were differentiated in the presence of DEHP for 2 days (for erythrocytes) or 3 days (for dendritic cells and neutrophils). Cells were then washed twice with PBS and resuspended at 1 × 105 cells in a 167 µL ATP detection sample buffer. Cell lysis and ATP quantitation were performed following the manufacturer’s protocol.
Kaiser L., Quint I., Csuk R., Jung M, & Deigner H.P. (2021). Lineage-Selective Disturbance of Early Human Hematopoietic Progenitor Cell Differentiation by the Commonly Used Plasticizer Di-2-ethylhexyl Phthalate via Reactive Oxygen Species: Fatty Acid Oxidation Makes the Difference. Cells, 10(10), 2703.
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7

Colorimetric and Luminescent Assays for ADP and ATP

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ADP and ATP production was determined using ADP Assay Kit (Colorimetric/Fluorometric) (ab83359, Cambridge, MA) and ATP Detection Assay Kit—Luminescence (No.700410, Cayman Chemical), respectively, according to the manufacture’s protocol. In brief, ATP levels are measured by luciferase/luciferin-mediated assay. For the measurement of ADP, ADP is converted to ATP and pyruvate. Then the generated pyruvate is quantified by the colorimetric method.
Xue W., Li X., Li W., Wang Y., Jiang C., Zhou L., Gao J., Yu Y., Shen Y, & Xu Q. (2022). Intracellular CYTL1, a novel tumor suppressor, stabilizes NDUFV1 to inhibit metabolic reprogramming in breast cancer. Signal Transduction and Targeted Therapy, 7, 35.
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8

Mitochondrial Function and Integrity in ARPE-19 Cells

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Mitochondrial function and integrity of ARPE-19 cells were evaluated using ATP Detection Assay Kit-Luminescence (Cayman) and JC-1 staining (Cayman), respectively. In the ATP detection, the luciferin in assay reagent was converted to oxyluciferin with light emission that was quantitatively proportional to the amount of intracellular ATP (n = 6 per treatment condition). JC-1 staining (1 μM 30 min, n = 6 per treatment condition) was performed as per the manufacturer's protocol. Both luminescence and fluorescence were detected using the Hidex Sense Microplate Reader (Hidex).
Lee J.J., Chang-Chien G.P., Lin S., Hsiao Y.T., Ke M.C., Chen A, & Lin T.K. (2022). 5-Lipoxygenase Inhibition Protects Retinal Pigment Epithelium from Sodium Iodate-Induced Ferroptosis and Prevents Retinal Degeneration. Oxidative Medicine and Cellular Longevity, 2022, 1792894.
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9

Astrocyte ATP and Glutamate Kinetics

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For the assessment of ATP and Glu release from astrocytes in the function of time, the cells were seeded onto a 96-well plate to a final density of 1.5 × 104 cells/well for 24 h. Afterwards, the cells were washed and supplied with fresh experimental medium (in case of ATP assessment) or HEPES-buffered solution (HBS; 120 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1.5 mM CaCl2, 10 mM glucose, 25 mM HEPES; pH = 7.4; in case of Glu assessment). After the incubations lasting 2, 5, 10, 15, or 30 min, the supernatants were collected and subjected for the measurands evaluation. For ATP, the luminescence-based test “ATP Detection Assay Kit—Luminescence” (Cayman; Ann Arbor, MI, USA; cat no. 700410) was applied according to the protocol recommended by the producer, and luminescence was assessed with the use of a Synergy H1 plate reader (Biotek; Winooski, VT, USA). For Glu measurement, the fluorescence-based test “Glutamate Assay Kit (Fluorometric)” (Abcam; Cambridge, UK; cat no. ab138883) was used according to the protocol recommended by the producer, and fluorescence (excitation λ = 550 nm, emission λ = 590 nm) was assessed with the use of a Victor X4 multilabel plate reader (PerkinElmer; Waltham, MA, USA).
Karbownik M.S., Sokołowska P, & Kowalczyk E. (2023). Gut Microbiota Metabolites Differentially Release Gliotransmitters from the Cultured Human Astrocytes: A Preliminary Report. International Journal of Molecular Sciences, 24(7), 6617.
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10

Murine CD8+ T Cell Isolation and Metabolic Profiling

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EasySep Mouse naïve CD8 T cell isolation kits (Stemcell Technologies cat:19858A); Mojosort mouse anti-APC nanobeads (Biolegend Cat:480072); ATP detection assay kit-luminescence (Cayman Chemical cat:700410); DAPI (Thermo Fisher cat:D1306); Seahorse XF Cell Mito Stress Test Kit (Seahorse Agilent cat:103015-100); 2-DG (Cayman Chemical cat:14325); SIINFEKL peptide (New England peptide Lot:V1355-37/40); recombinant human IL-2 (TECIN cat:Ro23-6019); recombinant murine IL-15 (PeproTech cat:210-15); poly-D-lysine (Millipore Sigma cat:P6407); Glutaraldehyde (Electron Microscopy Science cat:16000); NaBH4 (Alfa Aesar stock#:35788); Triton X-100 (PerkinElmer cat:N9300260).
Sun Y., Preiss N.K., Valenteros K.B., Kamal Y., Usherwood Y.K., Frost H.R, & Usherwood E.J. (2020). Zbtb20 restrains CD8 T cell immunometabolism and restricts memory differentiation and anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 205(10), 2649-2666.
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