ATP Bioluminescence Assay Kit HS II Chemische Eigenschaften,Einsatz,Produktion Methoden
Verwenden
Highly sensitive and quantitative determination of ATP. Contains a cell lysis reagent and can be applied for the detection of microbial contamination. The kit is well suited for use in tube luminometers and microplate-format luminometers. The cell lysis reagent exhibits good lysis efficiency with respect to a variety of eukaryotic and prokaryotic cells.
For a time constant light signal, use the ATP Bioluminescence Assay Kit CLS II.
For maximum sensitivity, the sample ATP must be in a minimum volume, and the luciferase reagent must not be diluted.
Allgemeine Beschreibung
Living things require a continual input of free energy for three major purposes: the performance of mechanical work in muscle contraction and other cellular movements, the active transport of molecules and ions, and the synthesis of macromolecules and other biomolecules from simple precursors.
The free energy used in these processes is derived from the surroundings and maintains an organism in a state that is far from equilibrium. In most processes, this special carrier of free energy is adenosine triphosphate (ATP), an energy-rich molecule. Its triphosphate unit contains two phosphoanhydride bonds. The turnover of ATP is very high. Motion, active transport, signal amplification, and biosynthesis (as is needed for cell proliferation) can occur only if ATP is continuously regenerated from ADP. Therefore, measurement of ATP can serve as a marker for cell proliferation.
The determination of ATP using bioluminescence is a well established technique. It uses the ATP dependency of the light-emitting, luciferase-catalyzed oxidation of luciferin for the measurement of extremely low concentrations of ATP.
Biochem/physiol Actions
The luciferase from Photinus pyralis (American firefly) catalyzes the following reaction:ATP + D-luciferin + O2? oxyluciferin + PPi + AMP + CO2 + light.The quantum yield for this reaction is about 90%. The resulting green light has an emission maximum at 562 nm.The Michaelis equation has the following form:light intensity = (Vmax x CATP)/(Km + CATP).At low ATP concentrations (CATPm), the formula is simplified to light intensity = Vmax x CATP/Km.From this equation, it becomes obvious that the light output is directly proportional to the ATP concentration (CATP), and is dependent on the amount of luciferase (Vmax) present in the assay.
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