Luminescent ATP Detection Assay Kit

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Functional Studies (Act) - Luminescent ATP Detection Assay Kit (AB113849)

Simultaneous quantification of mitochondrial respiration and glycolytic flux.

Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 μM] and FCCP [2.5 μM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).

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Functional Studies (Act) - Luminescent ATP Detection Assay Kit (AB113849)

Example of ATP standard curve using an opaque white plate.

The ATP standard curve was prepared as described in the protocol. Background-subtracted data values (mean +/- SD) are graphed.

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Functional Studies - Luminescent ATP Detection Assay Kit (AB113849)

Total cellular ATP concentration. ATP in SH-SY5Y cells cultivated at 21% and 5% O2 24 h after treatment with Aβ peptide and/or 18 h X-ray irradiation, normalized to cell count, and compared to respective controls. ATP concentration was about 1.3- to 1.8-fold higher at all conditions in cells cultivated at 5% O2 compared to 21% O2. Combination of Aβ peptide treatment and irradiation resulted in a significantly increased (~1.5-fold) ATP concentration at 5% O2 compared to the control. Samples were measured at least in duplicates (n = 2–4) in three independent experiments (N = 3). Mean ± SEM analyzed by two-way ANOVA with Tukey's multiple comparison test with p < 0.05 considered as significant. (∗∗ p < 0.01).

Dzinic, Tamara and Norbert A Dencher., Oxidative medicine and cellular longevity-vol. 2018 7567959., Fig 6, doi:10.1155/2018/7567959

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Functional Studies - Luminescent ATP Detection Assay Kit (AB113849)

Extracellular detection of ATP.

Analysis of the release of ATP by connexin hemichannels in stem cells using ATP luminescence kit (ab113849).

Cells were cultured in HBSS to induce hemichannel opening. Calcium and GAP-inhibitor were used to trigger hemichannel closure.
After two hours the supernatant was collected and ATP was measured according to the protocol (detergent was also applied).

Calcium treatment and inhibition by GAP decreased ATP concentration, compared to HBSS control. Graph shows data of three independent experiments.

This image is courtesy of an Abreview submitted by Heiko Lemcke.

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Functional Studies - Luminescent ATP Detection Assay Kit (AB113849)

ab113849 ATP detection kit cytotoxicity data. 25000 HepG2 cells were seeded into each well, allowed to adhere and treated for 4 hours with 25µM rotenone and vehicle control (DMSO) in glucose based complete media. After treatment, cells were lysed, exposed to the ATP substrate solution and signal was measured on a luminescent counter. Mean and standard deviation is plotted for 3 replicates from each condition. Rotenone induces cytotoxicity in HepG2 cells.