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Enliten atp assay system bioluminescence detection kit

Manufactured by Promega
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The ENLITEN ATP Assay System Bioluminescence Detection Kit is a laboratory equipment product designed to detect and quantify the presence of adenosine triphosphate (ATP) in biological samples. The kit utilizes bioluminescence technology to provide a sensitive and reliable method for ATP detection.

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Lab products found in correlation

Glomax 20 20 luminometer, by Promega (3 mentions) Glomax luminometer, by Promega (3 mentions) Paradigm i3, by Molecular Devices (3 mentions) Microplate reader, by Hidex (2 mentions) Dc protein assay, by Bio-Rad (2 mentions) M0187, by Greiner (2 mentions) Opteia human il 1β enzyme linked immunosorbent assay elisa kit 2, by BD (2 mentions) Fluostar optima instrument, by BMG Labtech (2 mentions) Sirna against cat 2, by OriGene (2 mentions) Clariostar, by BMG Labtech (2 mentions) Spectramax paradigm multi mode microplate reader, by Molecular Devices (2 mentions) 96 well solid white flat bottom polystyrene microplates, by Corning (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Mg 132 cas 133407 82 6, by Merck Group (1 mentions) Infinite m200 microplate analyzer, by Tecan (1 mentions) Bioluminescent somatic cell assay kit flasc, by Merck Group (1 mentions) Cary eclipse, by Agilent Technologies (1 mentions) Victor3, by Bio-Rad (1 mentions) Infinite 200 pro plate reader, by Tecan (1 mentions) Ff2000, by Promega (1 mentions)

Close spelling variants of this product

ENLITEN ATP Assay System (90 citations) ENLITEN ATP Assay (45 citations) ATP assay kit (35 citations) ENLITEN ATP Assay kit (28 citations) ATP bioluminescence assay kit (12 citations) ENLITEN (9 citations) ENLITEN-ATP kit (8 citations) ENLITEN kit (7 citations) ENLITEN® ATP Assay System Bioluminescence Detection Kit for ATP Measurement (7 citations) ENLITEN ATP assay system kit (6 citations)

61 protocols using enliten atp assay system bioluminescence detection kit

1

Ascorbate-mediated ATP Measurement

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Hypoxia inducible factor-1α inhibitor (CAS 934593-90-5) was purchased from Santa Cruz Biotechnology. MG-132 (CAS 133407-82-6) and cycloheximide (CAS 66-81-9) were purchased from EMD Millipore. L-Ascorbic acid was purchased from Macron Chemicals (Center Valley, PA). A stock solution of ascorbate (pH 7.0) was made under argon and stored in screw-top sealed test tubes at 4 °C. Ascorbate concentration was verified using, ε265 = 14,500 M−1 cm−123. The solution can be kept for several weeks without significant oxidation due to the lack of oxygen [23 (link)]. Intracellular ATP was measured using ENLITEN ATP assay system bio-luminescence detection kit from Promega (Madison, WI).
Wilkes J.G., O’Leary B.R., Du J., Klinger A.R., Sibenaller Z.A., Doskey C.M., Gibson-Corley K.N., Alexander M.S., Tsai S., Buettner G.R, & Cullen J.J. (2018). Pharmacologic ascorbate (P-AscH−) suppresses hypoxia-inducible Factor-1α (HIF-1α) in pancreatic adenocarcinoma. Clinical & experimental metastasis, 35(1-2), 37-51.
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2

Extracellular ATP Measurement Assay

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For the determination of the extracellular ATP, a luciferin–luciferase assay (ENLITEN ATP Assay System Bioluminescence Detection Kit, Promega) and a TECAN infinite M200 Microplate Analyzer (Lausanne, Switzerland) were used to record relative light units (RLU) as described by Liu, Sabirov, and Okada (2008) (link). As previously described (Xu et al., 2014 ), the assay was performed during 60 min in DMEM without serum or pH indicator at 37℃. The ecto-ATPase inhibitor ARL67156 was present in all experiments. Then, the medium was rapidly cooled to room temperature for the ATP assay and mixed with a 100 -μl aliquot of luciferin–luciferase reagent dissolved in dilution buffer provided with the assay kit, and chemiluminescence values (RLU) were recorded as an indication of ATP content. Even in the absence of any known ATP, considerable relative high RLU readings were caused by the medium or tissue. This appeared larger than the previously measured 1,200–1,600 RLU under control conditions (Xu et al., 2014 ), perhaps reflecting some effects of the reagents other than siRNA itself with which the cells had been treated.
Song D., Xu J., Bai Q., Cai L., Hertz L, & Peng L. (2014). Role of the Intracellular Nucleoside Transporter ENT3 in Transmitter and High K+ Stimulation of Astrocytic ATP Release Investigated Using siRNA Against ENT3. ASN NEURO, 6(4), 1759091414543439.
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3

Intracellular ATP Concentration Measurement

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To measure intracellular ATP concentrations, the ENLITEN ATP Assay System Bioluminescence Detection Kit (Promega) was used in accordance with the manufacturer’s instructions. Cultures of ATCC17978, Δadk, and Δdes6 were grown to the exponential phase and then treated with 64 μg/ml OA for 30 min. Next, the cells were harvested and then resuspended in 1% trichloroacetic acid (TCA) buffer. Prior to the assay, the samples were neutralized by diluting them fivefold with 250 mM Tris-acetate buffer (pH 7.75). The luminescence was measured using a microplate reader (Hidex).
Shin B, & Park W. (2015). Synergistic Effect of Oleanolic Acid on Aminoglycoside Antibiotics against Acinetobacter baumannii. PLoS ONE, 10(9), e0137751.
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4

Intracellular ATP Quantification in Hypoxia

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Cells were grown in 12-well plates as monocultures for 72 hours under normoxic (21% O2) or hypoxic (1.5% O2) conditions. Intracellular ATP was extracted by boiling water method as described previously by Yang et al. [34 (link)] with modification. Briefly, cells in 12-well plates were washed with cold PBS twice followed by adding 1 ml of boiling water to the wells directly. After repeated pipetting, cell extracts were cleared by centrifugation at 12000g for 5 min at 4°C. ATP levels in the supernatants were measured using the ENLITEN ATP Assay System Bioluminescence Detection Kit (Promega) following the manufacturer’s instructions. Total protein concentration was determined using the DC Protein Assay (BioRad) following the manufacturer’s recommended protocol.
Penzo-Méndez A.I., Chen Y.J., Li J., Witze E.S, & Stanger B.Z. (2015). Spontaneous Cell Competition in Immortalized Mammalian Cell Lines. PLoS ONE, 10(7), e0132437.
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5

ATP Quantification in L02 Cells

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The concentration of ATP in the L02 cells belonging to the different groups was measured using an ENLITEN ATP assay system bioluminescence detection kit (Promega, USA) following the manufacturer’s recommended protocol.
Zheng J., Chen L., Lu T., Zhang Y., Sui X., Li Y., Huang X., He L., Cai J., Zhou C., Liang J., Chen G., Yao J, & Yang Y. (2020). MSCs ameliorate hepatocellular apoptosis mediated by PINK1-dependent mitophagy in liver ischemia/reperfusion injury through AMPKα activation. Cell Death & Disease, 11(4), 256.
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6

Measuring ATP Levels in Mouse Oocytes

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Denuded oocytes (10 pooled/group) collected from unprimed or gonadotropin-stimulated mice were collected in 50 μl filtered ultrapure water and stored at −80 °C until use. To prepare standards, 10−7 M ATP standard stock was obtained from ENLITEN® ATP Assay System Bioluminescence Detection Kit (Promega) and was diluted with filtered ultrapure water. ATP levels were assayed using the Adenosine 5¢-triphosphate (ATP) bioluminescent somatic cell assay kit (FLASC, Sigma) according to the manufacturer’s instructions. Briefly, 100 μl ATP Assay Mix Working Solution was added to a 96-well plate (reaction vial) (M0187, Greiner) and allowed to stand at room temperature for 3 minutes. Somatic Cell ATP Releasing Reagent (100 μl), 50 μl filtered ultrapure water and 50 μl sample were added to a new tube and 100 μl was transferred to the reaction vial. ATP concentration was measured immediately using a luminometer (BMG, Clariostar, 76G58). Data are expressed as ATP content relative to the mean of controls or pmol/oocyte. Experiments were repeated a minimum of three separate times.
Wang Q., Stringer J.M., Liu J, & Hutt K.J. (2019). Evaluation of mitochondria in oocytes following γ-irradiation. Scientific Reports, 9, 19941.
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7

Kidney ATP Quantification Protocol

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Snap-frozen kidney tissue (n=3–4 animals/group) was homogenized in ice-cold 2% trichloroacetic acid. The supernatants were collected and neutralized with Tris-acetate (100 mM) and EDTA (2 mM, pH, 7.8). ATP concentrations were determined using the Enliten ATP Assay System Bioluminescence Detection Kit (Promega, Madison, WI).
Ding M., Tolbert E., Birkenbach M., Gohh R., Akhlaghi F, & Ghonem N.S. (2021). Treprostinil reduces mitochondrial injury during rat renal ischemia-reperfusion injury. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 141, 111912.
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8

Urinary Biomarkers Measurement Protocols

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The urinary (cell-free) concentrations of the candidate biomarkers were measured using ENLITEN® ATP Assay System Bioluminescence Detection Kit (FF2000, Promega, UK); Amplex® Red Acetylcholine/Acetylcholinesterase assay (Invitrogen™ Molecular Probes™, A12217, UK); Sievers Nitric Oxide Analyser (NOA™ 280i, Analytix, UK); BD OptEIATM human MCP-1 enzyme-linked immunosorbent assay (ELISA) (559017, BD biosciences, UK); Quantikine® human IL-5 ELISA Kit (R&D Systems®, D5000B, UK) and the OptEIATM Human IL-5 ELISA Set (555202, BD biosciences, UK) according to the manufacturers’ instructions.
Firouzmand S., Ajori L, & Young J.S. (2020). New participant stratification and combination of urinary biomarkers and confounders could improve diagnostic accuracy for overactive bladder. Scientific Reports, 10, 3085.
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9

Extracellular ATP Levels and IL-1β in Islet Cell Death

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Extracellular ATP levels, an indicator for human islet cell death, was evaluated under normal culture conditions as well as the high-glucose challenge conditions. High-glucose challenge was performed once a week as follows: media was collected and cultured cells were washed once with RPMI medium. Media was then replaced with high-glucose (20 mM) RPMI 1640 for 30 min. The media was then collected and stored at −80°C for analysis of extracellular ATP levels and IL-1β content. ATP levels in cell culture medium were analyzed with the ENLITEN® ATP Assay System Bioluminescence Detection Kit (Promega) according to the protocol, and luminescence was measured on a Fluostar Optima instrument (BMG Labtechnologies, Germany). IL-1β content was analyzed with the BD OptEIA human IL-1β enzyme-linked immunosorbent assay (ELISA) kit II according to the protocol (BD Biosciences Pharmingen, San Diego, CA). IL-1β was determined by measuring the optical density at 450 nm in a Labsystem Multiscan Plus fluorescence spectrophotometer. IL-1β and insulin concentration tested on the same time point were correlated in order to identify the relation between these two parameters.
Luo L.G, & Luo J.Z. (2015). Anti-apoptotic Effects of Bone Marrow on Human Islets: A Preliminary Report. Journal of stem cell research & therapy, 5(4), 274.
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10

ATP Quantification in 1C11 Precursors and PrP^null-1C11

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ATP content in 1C11 precursors cells and PrPnull-1C11 cells (2.5 x 105 cells) grown for 4 days in DMEM-Glutamax-FCS containing 25 mM glucose was assessed using the Enliten ATP Assay System Bioluminescence Detection Kit according to the manufacturer’s instructions (Promega, USA). The luminescence was recorded at λem = 560 nm (integrate period = 10 sec, slit width = 5 nm) using a Cary Eclipse (Varian Inc., Agilent Technology).
Arnould H., Baudouin V., Baudry A., Ribeiro L.W., Ardila-Osorio H., Pietri M., Caradeuc C., Soultawi C., Williams D., Alvarez M., Crozet C., Djouadi F., Laforge M., Bertho G., Kellermann O., Launay J.M., Schmitt-Ulms G, & Schneider B. (2021). Loss of prion protein control of glucose metabolism promotes neurodegeneration in model of prion diseases. PLoS Pathogens, 17(10), e1009991.
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