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咨询The Firefly Luciferase ATP Assay Kit allows cell viability to be measured using luminescence. This assay kit detects and quantifies ATP, an indicator of cell viability, by measuring assay readings in relative light units (RLUs) using a luminometer. This is a highly sensitive, highly stable kit with fast results that produces a stable, glowing signal proportional to the viable cell count. The Firefly Luciferase Reaction Mixture is made up of Firefly Luciferase and D-Luciferin.
Product Usage Information
Substrate Reagent Preparation:1. Thaw buffer; equilibrate supplied reagents at room temperature.2. Reconstitute Firefly Luciferase Reaction Mixture with entire bottle of Cell Lysis Buffer.3. Mix by gentle inverting. Directions for Use: Plate cells of interest at specific predetermined cell densities, at 100 μL per well, into the 96-well opaque assay plate. Prepare control wells containing media without cells. It is recommended to run samples in duplicates. Treat cells with compounds of interest and incubate according to desired specifications. Add 100 μL of Firefly Luciferase Reaction Mixture to each well containing cells and media only control wells. Mix contents on an orbital shaker for 2 min, then place on benchtop. Allow the plate to incubate at room temperature for an additional 15 min to stabilize luminescent signal. Obtain results from a plate reader with a Luminometer setting.Note: Absolute signal may vary by plate reader. Additional Components Not Supplied:· White opaque 96-well assay plate (i.e., White MaxiSorp/FluoroNunc 96-well plate, ThermoFisher Scientific #437591)· Single and multichannel pipettes· Orbital plate shaker· Plate reader with Luminometer setting.
Specificity/Sensitivity
All components in this kit are stable for at least 24 months when stored at -80ºC. After reconstitution of the Firefly Luciferase Reaction Mixture, aliquot into single use aliquots to avoid multiple freeze/thaw cycles. Store at 4ºC for up to a week or -20ºC for up to one month. Protect from light.
参考图片
Jurkat cells were plated at varying densities in a 96-well opaque assay plate. Firefly Luciferase Reaction Mixture was added to each well and incubated at room temperature for 15 min. Relative light units (RLUs) were determined by a plate reader, as shown in the figure.
An equal volume (100 μL) of Firefly Luciferase Reaction Mixture was added to either 50,000 Jurkat cells in 100 μL of cell culture medium or to medium only control wells. The luminescent signal was monitored for 5 different time intervals on a luminometer, as shown in the figure.