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ATP bioluminescence assays measure the level of ATP within the cell by:
- lysing the cell sample
- adding luciferase enzyme and luciferin
- measuring emitted light using a tube or microplate-based luminometer.
Luminescent ATP Detection Assay Kit (ab113849) irreversibly inactivates ATP degrading enzymes (ATPases) during the lysis step, ensuring that the luminescent signal obtained truly corresponds to the endogenous levels of ATP.
Total levels of cellular ATP can be used to assess cell viability, cell proliferation and cytotoxicity of a wide range of compounds and biological response modifiers.
|Detergent||1 x 20ml|
|Lyophilized ATP standard||1 vial|
|Lyophilized substrate||3 vials|
|Substrate Buffer||1 x 20ml|
The ATP standard curve was prepared as described in the protocol. Background-subtracted data values (mean +/- SD) are graphed.
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)).
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.
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