ATP Assay Kit

Sample

Cell culture supernatants, Tissue lysates, Cell lysates, Serum, plasma, saliva, or urine

Intended Use

The ATP Assay Kit is a simple fluorometric assay that measures the amount of total ATP present in biological samples in a 96-well microtiter plate format. Each kit provides sufficient reagents to perform up to 100 assays*, including blanks, ATP standards, and unknown samples. Sample ATP concentrations are determined by comparison with a known ATP standard. The kit has a detection sensitivity limit of 1.56 μM ATP.

Contents of Kit

Box 1 (shipped on blue ice packs)
1. ATP Standard: One 50 μL tube at 10 mM.
2. Glycerol Kinase: One 330 μL tube at 15.2 U/mL.
3. Glycerol-3-Phosphate Oxidase: One 610 μL tube at 37 U/mL.
4. Fluorometric Probe: One 50 μL tube in DMSO.
5. HRP: One 10 μL tube of a 100 U/mL solution in glycerol.
Box 2 (shipped on blue ice packs)
1. Neutralization Buffer: One 25 mL bottle.
2. 10× Assay Buffer: One 25 mL bottle.

Storage

Store the 10× Assay Buffer and Neutralization Buffer at room temperature. Store the Glycerokinase and the Glycerol-3-Phosphate Oxidase at -80°C. Store all other components at -20°C. The Fluorometric Probe is light sensitive and must be stored accordingly. Avoid multiple freeze/thaw cycles.

General Description

Adenosine triphosphate (ATP) is an organic molecule that provides energy for various cell processes such as enzymatic reactions leading to nerve impulses, muscle contraction, and synthesis of chemicals. ATP is found in all life forms and is classified as a nucleoside triphosphate consisting of three components: a nitrogenous base (adenine), the sugar ribose, and the triphosphate group. It is produced in animals from serial enzymatic reactions such as glycolysis or oxidative phosphorylation. In plants, sunlight is converted into ATP through a process known as photosynthesis. When used up metabolically, ATP is converted either to adenosine diphosphate (ADP) or to adenosine monophosphate (AMP). Other enzymatic reactions occur to regenerate ATP (which is also a precursor to DNA and RNA) and additionally ATP is used as a coenzyme.

Standard Curve

The following figures demonstrate typical ATP Assay Kit results. One should use the data below for reference only. This data should not be used to interpret or calculate actual sample results.

Figure 2: ATP Standard Curve.

Figure 3: ATP Detection in 293 and A549 cells using the ATP Assay Kit. Cell lysates were prepared according to the preparation of samples section above.