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Reagents & Solutions 800-356-9526 608-274-4330 enotes@promega.com
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FAQspeakCan I use a fluorometer rather than a luminometer to measure my luciferase activity? Can I use a liquid scintillation counter?We do not recommend using a fluorometer to measure luciferase activity for several reasons. First, firefly luciferase emits photons over a wide range of wavelengths, and a fluorometer is designed to detect energy over a fairly narrow window of wavelengths. Much of the light emitted by luciferase would not be detected by a fluorometer. Finally it is essential to turn off the excitation lamp; this can be difficult for many fluorometers. A liquid scintillation counter can be used. However, the sensitivity of detection will be approximately 100-fold less, and you will not be able to measure activity over several orders of magnitude of enzyme concentration. If using a liquid scintillation counter, measure activity in your sample by placing the sample in a microcentrifuge tube, then place the tube in the scintillation vial. Do not add scintillant; it will inactivate the luciferase. Ideally, the coincidence circuit of the scintillation counter should be turned off. If the circuit cannot be turned off, the luciferase concentration is proportional to the square root of the cpms. The GloMax™ Integrated Luminescence System provides superior instrumentation with built-in protocols to be used with all of our bioluminescent assays. Products may be covered by pending or issued patents or may have certain limitations on use. Please visit our patent and trademark web page for more information. |
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