Atplite assay kit

Manufactured by PerkinElmer

Sourced in United States, France, United Kingdom

The ATPlite assay kit is a luminescence-based detection system for quantifying the levels of adenosine triphosphate (ATP) in biological samples. The kit provides a sensitive and reliable method for measuring ATP, which is a fundamental indicator of metabolic activity and cell viability.

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Lab products found in correlation

Renilla luciferase assay system, by Promega (4 mentions) Prism 5, by GraphPad (4 mentions) Lb960, by Berthold Technologies (2 mentions) Empower 2 software, by Waters Corporation (2 mentions) Enspire multimode plate reader, by PerkinElmer (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Prism 6, by GraphPad (1 mentions) Recombinant cxcl12, by R&D Systems (1 mentions) Amd3100, by Merck Group (1 mentions) Tmre mitochondrial membrane potential assay kit, by Abcam (1 mentions) Nad nadh glo assay kit, by Promega (1 mentions) Ros glo assay kit, by Promega (1 mentions) Glucose uptake glo assay kit, by Promega (1 mentions) Lactate glo assay kit, by Promega (1 mentions) Topcount, by PerkinElmer (1 mentions) 96 well plate, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Fujifilm (1 mentions) Mso 1, by ZenBio (1 mentions) Hut smc, by PromoCell (1 mentions)

45 protocols using atplite assay kit

Intracellular ATP Quantification in Adipocytes

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Differentiated primary preadipocytes and 3T3‐L1 were lysed and intracellular ATP content measured by the ATP‐lite assay kit (Perkin Elmer, Villebon‐sur‐Yvette, France) (Braud et al., 2018 (link)).

Pini M., Czibik G., Sawaki D., Mezdari Z., Braud L., Delmont T., Mercedes R., Martel C., Buron N., Marcelin G., Borgne‐Sanchez A., Foresti R., Motterlini R., Henegar C, & Derumeaux G. (2021). Adipose tissue senescence is mediated by increased ATP content after a short‐term high‐fat diet exposure. Aging Cell, 20(8), e13421.

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Cytotoxicity Assay of Antiviral Compounds

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Cytotoxicity assays were carried out using the ATPlite^™ assay kit (PerkinElmer) following the manufacturer’s protocol. In short, in a white flat bottom 96-well plate, 25,000 MDCK cells were seeded and compounds and control (oseltamivir phosphate) were serial diluted (3x) and added to the well with final concentrations ranging from 250 μM to 38 nM. After a 24-hour incubation period, 10 μl of ATPlite solution was added to each well. The fluorescence reading was recorded using VictorTM X5, 2030 multilabel reader (PerkinElmer) and the CC₅₀ values were plotted using GraphPad PRISM 6.

Bottini A., De S.K., Wu B., Tang C., Varani G, & Pellecchia M. (2015). Targeting Influenza A virus RNA Promoter. Chemical biology & drug design, 86(4), 663-673.

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CXCR4/CXCL12 Regulation of SSC Expansion

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To test whether CXCR4/CXCL12 regulates in SSC expansion, equal concentration of SSC germ cell culture was plated on STO feeder and treated with the specific CXCR4 inhibitor, AMD3100 (1.25 μM; Sigma), or recombinant CXCL12 (10 ng/mL; R&D Systems), or both. Germ cells were cultured for 7 days after which they were gently dissociated from STO feeders and counted using a hemocytometer. In a repeat experiment, cell viability was assessed using the ATPlite assay kit per manufacturer (Perkin Elmer). In brief, 1 × 10⁴ germ cells were plated onto 96-well plates in defined culture media and treated with AMD3100, or the AKT inhibitor, LY29004 (33 μM). At day 4 and day 6 after treatment, cell viability was assessed by quantifying ATP-dependent luciferase activity.

Niu Z., Goodyear S.M., Avarbock M.R, & Brinster R.L. (2016). Chemokine (C-X-C) Ligand 12 Facilitates Trafficking of Donor Spermatogonial Stem Cells. Stem Cells International, 2016, 5796305.

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Metabolic Profiling of Cell Cultures

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MMP was investigated using the TMRE Mitochondrial Membrane Potential Assay Kit (Abcam). Intracellular ATP level at steady state was measured using ATPlite Assay Kit (PerkinElmer). Lactate level was measured using Lactate-Glo Assay Kit (Promega). Glucose uptake was assessed using Glucose Uptake-Glo Assay Kit (Promega). NAD⁺ and NADH levels were measured using NAD/NADH-Glo Assay Kit (Promega). ROS level was measured using ROS-Glo Assay Kit (Promega). The assays were performed according to the manufacturer's instructions. See supplemental information for detailed procedures.

Okarmus J., Havelund J.F., Ryding M., Schmidt S.I., Bogetofte H., Heon-Roberts R., Wade-Martins R., Cowley S.A., Ryan B.J., Færgeman N.J., Hyttel P, & Meyer M. (2021). Identification of bioactive metabolites in human iPSC-derived dopaminergic neurons with PARK2 mutation: Altered mitochondrial and energy metabolism. Stem Cell Reports, 16(6), 1510-1526.

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Cytotoxicity and Synergy Assessment

Cited 2 times

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Cytotoxicity was evaluated by tetrazolium-based 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) kit (Promega Corporation, Madison, WI, USA) as described previously [22 (link)]. The cell viability in terms of cellular ATP content was evaluated by an ATP-lite assay kit (PerkinElmer Life Sciences, Boston, MA, USA) [23 (link)]. The drug interaction for the UVC/WFA combined treatment was analyzed as previously described [24 (link)]. In brief, the formula for determining the synergy (α), i.e., additive, synergistic, or antagonistic for α = 1, >1, and <1, respectively, was listed as follows: α = survival fraction (SF) for UVC alone treatment × SF for WFA alone treatment/SF for UVC/WFA combined treatment. Cells were photographed at 100× magnification for morphology analysis.

Peng S.Y., Wang Y.Y., Lan T.H., Lin L.C., Yuan S.S., Tang J.Y, & Chang H.W. (2020). Low Dose Combined Treatment with Ultraviolet-C and Withaferin a Enhances Selective Killing of Oral Cancer Cells. Antioxidants, 9(11), 1120.

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Cytotoxicity Evaluation of Decellularized SFT

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Samples from each region of the decellularized SFTs (toe, middle, and ankle) were macerated and incubated in DMEM (300 mg tissue in 3 mL DMEM) for 72 h at 37°C with agitation. BHK (1 × 10⁵) or 3T3 (2.5 × 10⁵) cells in 200 μL of appropriate culture medium were seeded into the wells of 96-well plates (Nunc) and cultured for 24 h at 37°C in 5% (v/v) CO₂ in air. The culture medium was aspirated from the wells and 100 μL of the tissue extract plus 100 μL of appropriate culture medium (with double strength supplements) was added. The positive control was 40% (v/v) dimethyl sulfoxide (DMSO; Sigma) in culture medium (200 μL) and cells cultured in medium alone served as negative controls for cytotoxicity. The cells were cultured for 48 h before assessing cell viability by measuring cellular ATP. ATP was measured using the ATPLite™ assay kit (PerkinElmer) following the manufacturer's instructions. The luminescence (counts per second) in each well was measured using a TopCount (C9902 Perkin Elmer) luminescence plate reader.

Jones G., Herbert A., Berry H., Edwards J.H., Fisher J, & Ingham E. (2017). Decellularization and Characterization of Porcine Superflexor Tendon: A Potential Anterior Cruciate Ligament Replacement. Tissue Engineering. Part A, 23(3-4), 124-134.

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Quantitative Lactate and ATP Assays

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According to the manufacturer's protocol, lactate assay kit (Perkin-Elmer Life Sciences) was used to measure the level of lactate. Specifically, the cell culture supernatant was collected, deproteinized with a 10 kDa MWCO spin filter, then diluted with lactate assay buffer, and incubated with lactate probe and lactate enzyme mix at room temperature for 30 min, then measured for absorbance at 570 nm. According to the manufacturer's protocol, cellular ATP contents were measured by using an ATPlite assay kit (Perkin-Elmer Life Sciences). Briefly, 100 μl of cell lysate and substrate solution were mixed. Then the plates were vortexed for 2 min at 700 rpm and incubated for 10 min. Luminescence was assayed with a luminescence analyzer (Berthold LB960).

Zhang H., Guo X., Feng X., Wang T., Hu Z., Que X., Tian Q., Zhu T., Guo G., Huang W, & Li X. (2016). MiRNA-543 promotes osteosarcoma cell proliferation and glycolysis by partially suppressing PRMT9 and stabilizing HIF-1α protein. Oncotarget, 8(2), 2342-2355.

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Antiviral Compound Screening in Huh7.5.1 Cells

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Huh7.5.1 cells
were plated in 96-well plates at 10⁴ cells/well and incubated
overnight. The cells were infected with HCV-Luc in the presence of
increasing concentrations of the compound of interest. The viral level
was measured 48 h after treatment using a Renilla luciferase assay system (Promega, Madison, WI, USA). ATP-based cell
viability assay was carried out in parallel to evaluate the cytotoxicity
with an ATPlite assay kit (PerkinElmer, Waltham, MA, USA). The concentration
values that led to 50% viral inhibition and cytotoxicity (EC₅₀ and CC₅₀) were calculated using the nonlinear regression
equation in GraphPad Prism 5.0 software (GraphPad Software Inc., La
Jolla, CA, USA).

He S., Xiao J., Dulcey A.E., Lin B., Rolt A., Hu Z., Hu X., Wang A.Q., Xu X., Southall N., Ferrer M., Zheng W., Liang T.J, & Marugan J.J. (2015). Discovery, Optimization, and Characterization of Novel Chlorcyclizine Derivatives for the Treatment of Hepatitis C Virus Infection. Journal of Medicinal Chemistry, 59(3), 841-853.

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Quantifying Intracellular ATP in HRGECs

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Intracellular ATP levels of HRGECs were quantified by using an ATPlite assay kit according to the manufacturer’s instructions (PerkinElmer, Akron, OH, USA). Briefly, following cell exposure to HG with or without pretreatment with an inhibitor, mammalian cell lysis solution was added to cell-containing 96-well microplates. After shaking for 5 min at RT, equal amount of substrate was added and mixed for another 5 min at RT. The luminescent intensities were monitored 10 min later to indicate the ATP levels and mitochondrial activities.

Chen W., Xiang H., Chen R., Yang J., Yang X., Zhou J., Liu H., Zhao S., Xiao J., Chen P., Chen A.F., Chen S, & Lu H. (2019). S1PR2 antagonist ameliorate high glucose-induced fission and dysfunction of mitochondria in HRGECs via regulating ROCK1. BMC Nephrology, 20, 135.

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Quantifying Cellular ATP Levels

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ATP content was measured using the ATP-lite assay kit (Perkin Elmer, 6016941). Mitotic arrested cells (2 × 10⁴) were seeded in black 96-well plates and incubated with/without drugs for the indicated durations. Cells were incubated with 50 μL lysis buffer for 5 min and shaken with 50 μL substrate solution for 5 min. Luminescent readings of ATP contents were performed using a Victor 3 Model 1420-012 Multi-label Microplate Reader (Perkin Elmer).

Park Y.Y., Ahn J.H., Cho M.G, & Lee J.H. (2018). ATP depletion during mitotic arrest induces mitotic slippage and APC/CCdh1-dependent cyclin B1 degradation. Experimental & Molecular Medicine, 50(4), 46.

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