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The ATP Bioluminescence Assay

Overview

In fireflies, the luciferase enzyme converts a compound called luciferin into oxyluciferin, and produces light or “luminescence” as a result. This reaction requires energy derived from ATP in order to proceed, so researchers have exploited the luciferase-luciferin interaction to gauge ATP levels in cells. Given ATP’s role as the cell’s currency of energy, the ATP bioluminescence assay can provide insight into cellular metabolism and overall cell health.

In this video, JoVE discusses cellular respiration, specifically reviewing how glucose metabolism results in ATP production. This is followed by principles behind the ATP bioluminescence assay and a generalized protocol for this technique. Finally, a survey of how researchers are currently using the ATP bioluminescence assay to evaluate cell viability in a variety of experimental conditions.

Procedure

The ATP bioluminescence assay is a common technique used to quantify ATP levels and detect living, metabolically active cells. ATP or adenosine triphosphate is the primary source of energy for all living organisms, and by “all” we mean ALL. At the cellular level, ATP is generated through a set of metabolic processes called cellular respiration.

Today, we’ll briefly discuss the pathways involved in cellular respiration. Next, we’ll introduce the principles behind the ATP bioluminescence assay,

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Tags
ATP Bioluminescence AssayQuantify ATP LevelsDetect Living CellsMetabolically Active CellsAdenosine TriphosphateCellular RespirationGlycolysis PathwayGlucose MetabolismPyruvateAcetyl coenzyme ATricarboxylic Acid CycleElectron Transport Chain

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0:00

Overview

0:57

Cellular Respiration

2:43

Principles Behind the ATP Bioluminescence Assay

4:09

General Protocol for Measuring ATP in Cells

6:04

Applications

7:56

Summary

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