Bioluminescent ATP Quantification Protocol

ATP content was measured by a commercial bioluminescent assay (ATP bioluminescent assay kit, Merck) as previously described [27 (link)]. After treatments, ATP was extracted by boiling samples for 1 min in a solution containing 100 mM TRIS, 4 mM EDTA, pH 7.75. Bioluminescence measurements were carried out on 100 μL of each sample mixed with 100 μL of luciferin-luciferase solution using a standard luminometer. ATP content was calculated using a standard curve obtained by serial dilution of 2 μM ATP standard solution.

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Nele V., Tedeschi V., Campani V., Ciancio R., Angelillo A., Graziano S.F., De Rosa G, & Secondo A. (2023). Cerium-Doped Self-Assembling Nanoparticles as a Novel Anti-Oxidant Delivery System Preserving Mitochondrial Function in Cortical Neurons Exposed to Ischemia-like Conditions. Antioxidants, 12(2), 358.

Publication 2023

ATP bioluminescent assay kit

After treatments Bioluminescence measurements Dilution Edta Luciferase Luciferin Tris

Corresponding Organization :

Other organizations : University of Naples Federico II

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Variable analysis

independent variables

Treatments

dependent variables

ATP content

control variables

Boiling samples for 1 min in a solution containing 100 mM TRIS, 4 mM EDTA, pH 7.75
Bioluminescence measurements carried out on 100 μL of each sample mixed with 100 μL of luciferin-luciferase solution using a standard luminometer
ATP content calculated using a standard curve obtained by serial dilution of 2 μM ATP standard solution

controls

Positive control: 2 μM ATP standard solution
Negative control: Not explicitly mentioned

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