Atp Assay Buffer
Atp Assay Buffer - Related Videos
08:32
The ATP Bioluminescence Assay
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2023
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...
08:05
Assaying Protein Kinase Activity with Radiolabeled ATP
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Cited by 15 •
2017
Protein kinases are highly evolved signaling enzymes and scaffolds that are critical for inter- and intracellular signal transduction. We present a protocol for measuring kinase activity through the use of radiolabeled adenosine triphosphate ([γ-32P] ATP), a reliable method to aid in elucidation of cellular signaling regulation.
02:56
An ATP Utilization Assay for Determining Bacterial Survival Under Antibiotics Treatment
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2025
This video describes the simple ATP-measuring assay using luciferase glow reagent. This assay is used to quantify Neisseria gonorrhoeae survival after treatment with ceftriaxone.
02:56
Buffers
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2020
A solution containing appreciable amounts of a weak conjugate acid-base pair is called a buffer solution, or a buffer. Buffer solutions resist a change in pH when small amounts of a strong acid or a strong base are added. A solution of acetic acid and sodium acetate is an example of a buffer that consists of a weak acid and its salt: CH3COOH (aq) + CH3COONa (aq). An example of a buffer that consists of a weak base and its salt is a solution of ammonia and ammonium chloride: NH3 (aq) + NH4Cl...
01:31
ATP Yield
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2019
Cellular respiration produces 30 - 32 ATP per glucose molecule. Although most of the ATP results from oxidative phosphorylation and the electron transport chain (ETC), 4 ATP are gained beforehand (2 from glycolysis and 2 from the citric acid cycle). The ETC is embedded in the inner mitochondrial membrane and is comprised of four main protein complexes and an ATP synthase. NADH and FADH2 pass electrons to these complexes, which pump protons into the intermembrane space. This distribution of...
FAQs
The buffer establishes the pH and ionic environment required for luciferase to convert ATP and luciferin into measurable light. If these chemical conditions vary, the reaction may produce a different signal even when ATP levels are unchanged. Maintaining suitable conditions therefore helps the emitted light reflect ATP concentration rather than uncontrolled differences in reaction chemistry.
These functions address different stages of measurement. Lysis releases intracellular ATP from neurons, glial cells, or other samples, while stabilization helps preserve the released molecule before and during detection. Supporting both steps reduces the risk that incomplete extraction or ATP loss will distort the measured signal and complicate comparisons among experimental samples.
Consistent composition improves assay sensitivity, reproducibility, and interpretation. Because the buffer controls conditions for extraction and the luciferase reaction, changes in its formulation can alter the measured light independently of biological ATP differences. Using the same composition across samples makes shifts in signal easier to attribute to cellular energy status or experimental treatment.
In the luciferase-catalyzed reaction, ATP and luciferin generate light, and the signal is proportional to ATP concentration under suitable assay conditions. The measurement can therefore indicate relative cellular energy status in neurons or glial cells. Interpreting differences requires attention to the shared buffer conditions, since chemical variation can influence signal intensity.
A typical workflow uses the prepared solution to help lyse the sample and extract ATP, then provides the chemical environment for the luciferase and luciferin reaction. The resulting light is measured and related to ATP concentration. In neuroscience experiments, this sequence supports comparison of ATP levels across neural samples exposed to different conditions or treatments.
It is useful when researchers need to examine cellular energy status, mitochondrial function, or viability in neurons and glial cells. Measurements can also reveal metabolic changes associated with injury, disease, or experimental treatments. By supporting consistent ATP extraction and detection, the buffer helps connect bioluminescent assay results with changes in neural-cell physiology.