Description
General description
ATP bioluminescence assay is a sophisticated micro count method. It works based on the capability of fireflies to generate its glowing cold light. Rise in proliferation for all cytohexdrin concentrations can be determined by ATP bioluminescence assay.
Application
The ATP Bioluminescence Assay Kit CLS II is optimized for easy use in tube luminometers and microplate-format luminometers. The kit exhibits a constant light signal that is sustained for several minutes. The kit is well suited for kinetic studies and ATP determinations in coupled enzymatic reactions.
For more highly sensitive ATP determinations, use the ATP Bioluminescence Assay Kit HS II.
ATP Bioluminescence Assay Kit CLS II has been used in the determination of the ATP synthetic activity of the H⁺-ATP synthase, ATP bioluminescent assay and intracellular measurement of ATP levels.
Biochem/physiol Actions
The ATP Bioluminescence Assay Kit CLS II is specially developed for applications in which constant light signals are required for kinetic studies of enzymes and metabolic studies, or if coupled enzymatic assays are applied. If ATP determinations are performed manually, the CLS Kit provides high reproducibility due to the constant signal generation.
The sensitivity of the kit is reduced by a factor of 10, compared to the ATP Bioluminescence Assay Kit HS II, which is recommended for determinations that are in the high-sensitivity range. The ATP Bioluminescence Assay Kit HS II also contains an efficient cell lysis reagent, and can be used for the detection of ATP in microorganisms or animal cells.
Packaging
1 kit containing 2 components.
Preparation Note
Working concentration: Working range: 10⁻⁶ to 10⁻¹¹ M ATP
Working solution: Preparation of working solutions
Luciferase Reagent
Dissolve the whole content of one bottle by carefully adding 10 ml of double-dist. water. Incubate for five minutes at 0 to 4 °C without stirring or shaking. Mix for a homogeneous solution by carefully rotating the bottle. Do not shake. The reagent is stable for one day at 15 to 25 °C or for one week when stored at 0 to 4 °C. However, set up a standard curve each day, because a slightly loss of light activity occurs during this time (approx. 20% after 5 days).
ATP Standard
Each bottle contains approx. 10 mg ATP (> 98% purity; Mr 605.2). The exact amount of ATP is determined individually for each lot as indicated on the label. Dissolve the content of one bottle by addition of the appropriate volume of double-dist. water to get a final concentration of 10 mg/ml or 16.5 mM, respectively (e.g., 960 μl to 9.60 mg ATP).
Roche recommends reconstituting Luciferase Reagent in water and not in buffer.
Other Notes
For life science research only. Not for use in diagnostic procedures.
