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Aposensor atp cell viability bioluminescence assay kit

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The ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit is a laboratory tool that measures cellular ATP levels. It utilizes a bioluminescent reaction to quantify ATP, which is an indicator of cell viability.

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Lab products found in correlation

Aposensor adp atp ratio bioluminescence assay kit, by Abcam (1 mentions) Cytation 5 luminometer, by Agilent Technologies (1 mentions) Glomax navigator, by Promega (1 mentions) Lactate colorimetric fluorometric assay kit, by Abcam (1 mentions) Deproteinizing sample preparation kit, by Abcam (1 mentions) Lumat 9507, by Berthold Technologies (1 mentions)

Spelling variants of this product

ATP Assay Kit (195 citations) ApoSENSOR cell viability assay kit (8 citations) ATP Bioluminescence Assay Kit (3 citations) ApoSENSOR assay kit (3 citations) ApoSENSOR ATP Bioluminescence Assay Kit (2 citations) Cell Viability Kit (2 citations)

7 protocols using aposensor atp cell viability bioluminescence assay kit

1

ATP and ADP Bioluminescence Assay

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The ApoSENSOR ATP Cell Viability Bioluminescence Assay Kit (BioVision, № K254 and ApoSENSOR ADP/ATP Ratio Bioluminescence Assay Kit (BioVision, № K255) were used to assess the status of both gonad cells. The analyses were conducted according to the manufacturer’s protocols. Determination of ATP concentration is based on the reaction of oxidative decarboxylation of luciferin catalyzed by luciferase, in the presence of high energy ATP and magnesium ions. The light intensity was measured at a wavelength of 562 nm. ADP level was measured by its conversion to ATP that is detected using the same reaction. Results were expressed as nmol ATP ∙ mg−1 protein. Protein content was measured according to Bradford34 (link), using bovine albumin (protein content > 95%, Fluka) as standard.
Poprawa I., Chajec Ł., Chachulska-Żymełka A., Wilczek G., Student S., Leśniewska M, & Rost-Roszkowska M. (2022). Ovaries and testes of Lithobius forficatus (Myriapoda, Chilopoda) react differently to the presence of cadmium in the environment. Scientific Reports, 12, 6705.
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2

ATP Depletion Effects on Cell Viability

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To study the effects of ATP depletion (Lobo G.P. ATP modulates PTEN subcellular localization in multiple cancer cell lines, Human molecular genetic 2008), cells were incubated with 37 °C for 4 h in serum-free and glucose-free RPMI 1640 medium containing 10% FBS and 1% penicillin-streptomycin, and sodium azide (NaN3), which affects mitochondrial phosphorylation and, subsequently, lower cellular content. The optimal concentration of NaN3 was determined by the significant cytotoxic effects of MX 300 μM on MCF-7/MX cells. The cells were then placed on ice, the medium removed, and the cell monolayers were washed twice in ice-cold glucose-free HBSS and ready to be used. An ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit (BioVision, Catalog number: K254-200) was used to analyze the expression of ATP.
Chang F.W., Fan H.C., Liu J.M., Fan T.P., Jing J., Yang C.L, & Hsu R.J. (2017). Estrogen Enhances the Expression of the Multidrug Transporter Gene ABCG2—Increasing Drug Resistance of Breast Cancer Cells through Estrogen Receptors. International Journal of Molecular Sciences, 18(1), 163.
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3

Quantifying Cellular ATP Levels

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ATP levels were measured by the ApoSENSOR ATP Cell Viability Bioluminescence Assay Kit (K254, BioVision) according to the manufacturer’s instructions. Brief, 10 μl of indicated cells (containing 5000 cells) was transferred into luminometer plate, followed by the addition of 100 μl of the Nucleotide Releasing Buffer. Then, 10 μl ATP Monitoring Enzyme was added to the cell lysate, and the plate was read within ~ 1-2 min in a Cytation 5 luminometer (BioTek Instruments, Inc) or a GloMax® Navigator (Promega).
Weng H., Huang F., Yu Z., Chen Z., Prince E., Kang Y., Zhou K., Li W., Hu J., Fu C., Aziz T., Li H., Li J., Yang Y., Han L., Zhang S., Ma Y., Sun M., Wu H., Zhang Z., Wunderlich M., Robinson S., Braas D., Hoeve J.T., Zhang B., Marcucci G., Mulloy J.C., Zhou K., Tao H.F., Deng X., Horne D., Wei M., Huang H, & Chen J. (2022). The m6A Reader IGF2BP2 Regulates Glutamine Metabolism and Represents a Therapeutic Target in Acute Myeloid Leukemia. Cancer cell, 40(12), 1566-1582.e10.
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4

Zygote ATP Quantification Using Bioluminescence

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The ATP content of 19 pools (3 zygotes each pool) per group (Control or Treated with 6 mM of BOHB, as described below) was measured using the ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit (BioVision, Milpitas, CA, USA, cat. # k-254) according to the manufacturer’s instructions. Briefly, a mix containing 10 μl of “ATP monitoring enzyme” and 90 μl of “Nucleotide releasing buffer” were prepared for each sample and added to a 96-well plate. The plate was read on a BMG Labtech Fluorstar Optima microplate reader (BMG Labtech, Ortenberg, Germany) to obtain the background luminescence values. Next, 50 μl of “Nucleotide releasing buffer” was mixed gently into each microtube containing the zygotes, and the contents were transferred to a 96-well plate. Samples were incubated for 2 min at RT and read to obtain the luminescence values. The background luminescence was subtracted from all readings. In parallel, a five-point ATP standard curve (0.15, 0.075, 0.0375, 0.01875, and 0.009375 μM) was generated and used to estimate the ATP concentration in each sample.
Sangalli J.R., Sampaio R.V., del Collado M., da Silveira J.C., De Bem T.H., Perecin F., Smith L.C, & Meirelles F.V. (2018). Metabolic gene expression and epigenetic effects of the ketone body β-hydroxybutyrate on H3K9ac in bovine cells, oocytes and embryos. Scientific Reports, 8, 13766.
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5

Quantifying Lactate and ATP Levels

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Lactate levels were determined with the Lactate Colorimetric/Fluorometric Assay Kit (K607, BioVision), and the colorimetric assay was performed per the manufacturer’s recommendation with an additional deproteinization step to remove LDH from the cells cultured with FBS. Briefly, 1 × 106 cells from each group were washed once with PBS, lysed with 160 μL Lactate Assay Buffer (provided in the kit) and centrifuged to remove the insoluble materials. 150 μL of the soluble fraction was then transferred to a new Eppendorf tube for deproteinization with Deproteinizing Sample Preparation Kit (ab204708, Abcam). The deproteinized samples were then added to a 96-well plate in triplicates with appropriate dilution; a series of lactate standards with different dilution factors were also added to the plate for standard curve preparation. After incubation with the reaction mix (containing lactate enzyme mix and probe) for 30 minutes, absorbance of the samples at 570nm was measured and the lactate levels were calculated using the standard curve prepared. ATP levels were examined by the ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit (K254, BioVision) in accordance with the supplier’s direction.
Qing Y., Dong L., Gao L., Li C., Li Y., Han L., Prince E., Tan B., Deng X., Wetzel C., Shen C., Gao M., Chen Z., Li W., Zhang B., Braas D., ten Hoeve J., Sanchez G.J., Chen H., Chan L.N., Chen C.W., Ann D., Jiang L., Müschen M., Marcucci G., Plas D.R., Li Z., Su R, & Chen J. (2021). R-2-hydroxyglutarate attenuates aerobic glycolysis in leukemia by targeting the FTO/m6A/PFKP/LDHB axis. Molecular cell, 81(5), 922-939.e9.
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6

Algae Viability Assay via ATP

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Intracellular ATP concentration was used to determine the viability of algae that had undergone chilling and freezing with an ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit (BioVision, U.S.A). For each sample, an aliquot of 10 μL of the thawed sample was transferred into a vial supplied with the kit and 100 μL of Nucleotide Releasing Buffer was added. The mixture was incubated for 3 min before the addition of 5 μL of ATP Monitoring Enzyme. The ATP concentration was measured using the light produced when the sample reacted with firefly luciferase using a Lumat 9507 luminometer (Berthold Technologies, Wildbad, Germany). The amount of light produced was directly proportional to the amount of ATP present in the sample. Dinoflagellates were then cultured, further assessing the survivability.
Chong G., Tsai S., Wang L.H., Huang C.Y, & Lin C. (2016). Cryopreservation of the gorgonian endosymbiont Symbiodinium. Scientific Reports, 6, 18816.
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7

Cellular ATP Quantification via Bioluminescence

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The cellular ATP levels were measured using ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit (Biovision #K254), according to manufacturer’s instructions. In brief, 10000 cells were lysed in buffer supplied with the product and assayed for ATP levels. The assay utilizes the enzyme luciferase to catalyze the formation of light from ATP and luciferin, where the light was measured using a luminometer.
Saini S.K., Mangalhara K.C., Prakasam G, & Bamezai R.N. (2017). DNA Methyltransferase1 (DNMT1) Isoform3 methylates mitochondrial genome and modulates its biology. Scientific Reports, 7, 1525.
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