CATALOG INDEX
BY ALPHABETICAL
ORDER

CATALOG INDEX
BY DEPARTMENT

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• Cell viability by intra-cellular ATP
• Measures both intra-cellular and extra-cellular ATP
• Optimized for eukaryotic cells
• Minimal sample preparation
• Fast: 50 samples can be processed in less than 1 hour.
• Each kit includes:
  • Lyophilized Luciferin/Luciferase reagent and diluent
  • Lyophilized ATP standard and diluent
  • Pre-aliquoted standard curve calibrator diluents
  • ATP extraction Reagent optimized for eukaryotic cells
  • FireZyme's proprietary ATP stabilizing solution
• Other optimized Cell Viability Kits are available for sperm or bacterial cells.

Principle of test

FireZyme Cell Culture Viability Test Kits use a bioluminescence reaction to determine the intracellular ATP content of a simple of eukaryotic cells. ATP from the prepared sample reacts with the firefly enzyme, luciferase, to oxidize the substrate luciferin, which generates light. The light output of the reaction is measured in a luminometer.

ATP is a useful biochemical indicator because it is the "curency" of energy exchange within a livin cell. Its concentration in viable cells is strictly regulated.

To measure ATP content, firefly luciferin-luciferase is added to the prepared sample. The firefly luciferase catalyzes the production of light by the following bioluminescence reaction:

ATP : Luciferin + O₂ =  (Luciferase) =
Oxyluciferin + AMP + PPI + CO₂ + LIGHT

Since Luciferase and Luciferin (LL) are in excess in the reaction mixture, light output is linearly related to ATP concentration over five orders of magnetude. LL light output is very sensitive to the ionic strength and pH of the medium; therefore the samples are diluted 20 fold in FireZime's proprietary Buffer-Diluent. This dilution step eliminates test variation between samples due to differences in growth media and supplements, media ionic strength, and possible inhibitors of the LL reaction.

Standard Curve

The FireZyme Cell Culture Cell Viability Test Kit was used to make a standard curve with ATP standards in the range of 4µg/mL to 0.5µg/mL of ATP.The RLUs Were obtained using a FireZyme Model 2000 Luminometer with a delay time of 1 second and an integration time of 5 seconds.

Cell Culture Growth Curve

A growth curve for a human lymphoma cell line (U937) in RPMI containing 10% FBS was determined using the FireZyme Cell Culture Viability Test Kit. Viable cell numbers were determined by Trypan Blue exclusion.

Correlation of Intracellular ATP with Cell Number

The amount of ATP per U937 cell was determined using the FireZyme Culture Cell Viability Test. As above, the number of viable cells was estimated by microscopic cell counting after staing with Trypan Blue.