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119107 Sigma-Aldrich

ATP Assay Kit

Synonym: Luciferin-Related Kit

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Properties

Quality Level   100
usage   sufficient for 200 tests
mfr. no.   Calbiochem®
storage condition   OK to freeze
  avoid repeated freeze/thaw cycles
storage conditions   = -70C
input   sample type sludge
sample type blood
sample type milk
sample type soil
sample type urine
detection method   chemiluminescent

Description

General description

Cell death (especially apoptosis) is an energy-dependent process that requires ATP. As ATP levels fall to a point where the cell can no longer perform basic metabolic functions, the cell will die. A typical apoptotic cell exhibits a significant decrease in ATP level. Therefore, loss of ATP level in the cell has been used as an indicator of cell death. In contrast, increased levels of ATP has been an indicator of cell proliferation. Calbiochem® brand ATP Assay Kit utilizes bioluminescent detection of the ATP levels for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or Beta Counter.

Figure 1: Calculations

The assay is based on the firefly luciferase-catalyzed oxidation of D-luciferin in the presence of ATP and oxygen, whereby the amount of ATP is quantified by the amount of light (hv) produced.

Packaging

1 kit in Glass bottle

Components

One vial of Luciferase ATP Monitoring Enzyme, Enzyme Reconstitution Buffer, 1 bottle of Nucleotide Releasing Reagent, one vial of ATP Standard, and a user protocol.

Warning

Toxicity: Multiple Toxicity Values, refer to MSDS (O)

Specifications

Assay Time: 1 h

Principle

The assay can be fully automatic for high throughput (10 s/sample) and is extremely sensitive (detects 10-100 mammalian cells/well). The high sensitivity of this assay has led to many other applications for detecting ATP production in various enzymatic reactions, as well as for detecting low level bacterial contamination in samples such as blood, milk, urine, soil, and sludge.

Preparation Note

Reagent Reconstitution1. Reconstitute ATP Monitoring Enzyme with 2ml of the Enzyme Reconstitution Buffer. Mix well by gentle pipetting. The reconstituted enzyme is stable for up to 2 months at 4°C.2. Prepare an ATP standard solution by dissolving the 1 mg ATP into 1 ml of H2O. The solution is stable for several weeks at -20°C.

Storage and Stability

Upon arrival store the entire contents of the kit at -70°C.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety & Documentation

Safety Information

Protocols & Articles
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