SureLight™ Glow – Firefly Luciferase Cell Viability Assay

$90.00

SKU: AD-LUC-100 Categories: ,

Description

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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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

Reagent Preparation:

  1. Thaw buffer; equilibrate to room temperature
  2. Reconstitute lyophilized enzyme/substrate with buffer
  3. Mix by vortexing or gently inverting

Protocol for Assay:

  1. Prepare opaque-walled multi-well plates with mammalian cells in culture medium, (e.g.,100µl per well for 96-well plates)
  2. Prepare control wells containing medium without cells
  3. Add the test compound to experimental wells and incubate according to the cell culture protocol
  4. Equilibrate the plate and its contents at room temperature, approximately 15-30 minutes
  5. 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)
  6. Mix contents on an orbital shaker to induce cell-lysis
  7. Allow the plate to incubate at room temperature for 5-15 minutes to stabilize luminescent signal
  8. Read results on luminometer

 

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Size:
10 mls/100 assays
Form / Storage:
For long term storage, both the lyophilized enzyme/substrate powder and cell-lysis buffer should be stored at -20°C. For short term storage, if partially used, the reconstituted reagent can be stored at 4°C for up to one week.
Stability:

 

 

 

Technical Info:

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 µl of RPMI-1640/10% FBS medium. 100 µl 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 µl) of cell viability reagent were added to either 50,000 THP-1 cells in 100 µl 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.