As an indicator of cellular energy status, ADP/ATP ratio was performed using the Abcam Bioluminescent ADP/ATP Ratio Assay Kit and performed according to manufacturer’s instructions (ab65313, Cambridge, MA, USA). In brief, 100 µL of ATP Reaction Mix (1× ATP Monitoring Enzyme in Nucleotide Releasing Buffer) was loaded on a 96-well plate, followed by a baseline plate reading. To measure ATP levels, 100 µg of cortical lysate was then added to each well, followed by 2 min incubation and plate read. Finally, to determine ADP levels, 100 µL ADP (1× ADP Converting Enzyme in Nucleotide Releasing Buffer) was then added to each well and incubated for 2 min, followed by a final plate reading. ADP and ATP content quantified using a standard curve. Bioluminescence measured on Varioskan LUX Plate Reader (Thermo Scientific, MA, USA).
Bioluminescent adp atp ratio assay kit
Manufactured by Abcam
Sourced in United States
The Bioluminescent ADP/ATP Ratio Assay Kit is a laboratory tool designed to measure the ratio of adenosine diphosphate (ADP) to adenosine triphosphate (ATP) in a sample. This kit utilizes a bioluminescent reaction to quantify the levels of these essential cellular energy molecules.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microplate reader, by Agilent Technologies (2 mentions)
Jc 1 fluorescence probe, by Thermo Fisher Scientific (2 mentions)
Ab65313, by Abcam (1 mentions)
Cytation 3 imaging reader, by Agilent Technologies (1 mentions)
Synergy h4 hybrid microplate reader, by Agilent Technologies (1 mentions)
6 protocols using bioluminescent adp atp ratio assay kit
1
Quantifying Cellular Energy Homeostasis
2
Erlotinib-Induced Mitochondrial Dysfunction
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Cells were seeded in 96-well plates and treated with 25 μM erlotinib for 24 h. Intracellular ADP/ATP ratio was measured using a bioluminescent ADP/ATP Ratio Assay Kit according to the manufacturer’s instructions (Abcam, Cambridge, MA). Mitochondrial membrane depolarization was determined using the JC-1 fluorescence probe according to the manufacturer’s instructions (Molecular Probes). Cells were labeled with 2 μM JC-1 for 30 min at 37 °C in the dark and then analyzed at 488 nm excitation with 530/30 or 585/42 nm bypass emission filters using a fluorescence microplate reader (Bio-TeK Instruments).
3
Erlotinib-Induced Mitochondrial Dysfunction
4
Niclosamide and ND-Nic Impact on ATP Levels
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A total of 150,000 cells per well were seeded onto a 6-well plate and left to sit for 24 h. Cells were treated with 2 µM niclosamide and 5 µM ND-Nic for 48 h. Cells were then harvested via trypsinisation and lysed in nucleotide releasing buffer (Abcam, USA ab65313). ATP levels were obtained using ADP/ATP Ratio Bioluminescent Assay Kit (Abcam, USA ab65313). Luminescence signal was measured using BioTek Cytation3 Imaging Reader.
5
ATP and ADP Quantification in Cortical Tissue
6
Measuring Astrocyte ATP Levels
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WT and Mut primary astrocytes were thawed and seeded at a density of 5 × 103 cells per well of a PDL pre-coated 96-well plate for 24 h incubation in ‘astrocyte medium’ followed by further 48 h incubation in DMEM-HG or DMEM-GS medium. The ADP/ATP Ratio Bioluminescent Assay Kit (#ab65313; Abcam) was used to measure ATP levels according to the manufacturer’s instructions, employing Veritas microplate luminometer (Turner BioSystems/Promega, Madison, WI, USA). ATP levels were normalized to biomass, obtained by Crystal Violet staining [24 (link)].
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