SureLight® Glow – Firefly Luciferase Cell Viability Assay
The SureLight® Glow firefly luciferase cell viability assay provides a homogeneous and highly sensitive method for quantifying ATP, an indicator of cell viability, in cell cultures. The assay generates a stable, glow-type luminescent signal directly proportional to the amount of viable cells present in the cell culture. SureLight® Glow meets or exceeds the specifications and performance of leading competitors at a greater than 40% cost savings.
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|Simple; mix, plate & read||yes|
|Stable, glow-type signal||yes|
|Fast results||5-15 minutes|
|Extended half-life||> 2 hours|
This kit includes:
- Firefly Luciferase (lyophilized)
- D-Luciferin (lyophilized)
- Lysis Buffer (frozen)
Material to be supplied by user:
- Opaque-walled multi-well plate
- Multichannel pipette
- Plate shaker
- Luminometer or imaging device capable of reading multi-well plates
- Thaw buffer; equilibrate to room temperature
- Reconstitute lyophilized enzyme/substrate with buffer
- Mix by vortexing or gently inverting
Protocol for Assay:
- Prepare opaque-walled multi-well plates with mammalian cells in culture medium, (e.g.,100?l per well for 96-well plates)
- Prepare control wells containing medium without cells
- Add the test compound to experimental wells and incubate according to the cell culture protocol
- Equilibrate the plate and its contents at room temperature, approximately 15-30 minutes
- Add a volume of cell viability reagent equal to the volume of medium containing cells in each well (e.g., 100ul of reagent to 100ul of cell culture medium)
- Mix contents on an orbital shaker to induce cell-lysis
- Allow the plate to incubate at room temperature for 5-15 minutes to stabilize luminescent signal
- Read results on luminometer
Correlation of the number of viable cells with the luminescent signal. Two-fold serial dilution of THP-1 cells (from 50,000 to 49 cells per well) was prepared in 100 ?l of RPMI-1640/10% FBS medium. 100 ?l of the reagent was added to each well and the reaction was incubated for 15 minutes at room temperature. The luminescent signal was measured on a luminometer and expressed in relative light units (RLU).
Luminescent signal stability over time. Equal amounts (100 ?l) of cell viability reagent were added to either 50,000 THP-1 cells in 100 ?l of RPMI1640/10% FBS medium (viable cells) or to medium alone (control; no cells). The luminescent signal was monitored every 10 minutes over the indicated period of time on a luminometer.
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