Ordering information
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FR201-01
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50 rxns
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|
FR201-02
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200 rxns
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Storage: at -20oC in dark for one year; reconstituted luciferase assay reagent (substrate in Reaction Buffer) stored in aliquots at -20oC in dark for one month or at -70oC for one year
Description
Trans DetectTM Double-Luciferase Reporter Assay Kit provides an efficient means of performing dual-reporter assays. In the assay, the activities of firefly (photinus pyralis) and renilla (sea pansy) luciferases are measured sequentially from a single sample. The firefly luciferase reporter is measured first by adding Luciferase Reaction Reagent to generate a stable luminescent signal. After quantifying the firefly luminescence, this reaction is quenched, and the renilla luciferase reaction is simultaneously initiated by adding Luciferase Reaction Reagent II to the same tube.
1.Firefly and renilla luciferases are used as reporters
2.high sensitivity
3.fast detection
4.broad range of detection
Kit Contents
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Component
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FR201-01 (50 rxns)
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FR201-02 (200 rxns)
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Luciferase Reaction Buffer
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5 ml
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20 ml
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Luciferase Reaction Substrate (Lyophilized)
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1 vial
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4 vials
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Luciferase Reaction Buffer II
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5 ml
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20 ml
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Luciferase Reaction Substrate II (50×)
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100 μl
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400 μl
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|
Cell Lysis Buffer (5×)
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5 ml
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20 ml
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Procedures
1. Prepare Luciferase Reaction Reagent
Dissolve the lyophilized Luciferase Reaction Substrate (whole vial) by adding 5 ml of Luciferase Reaction Buffer to the vial. The prepared Luciferase Reaction Reagent should be stored in the dark.
2. Prepare Luciferase Reaction Reagent II
One volume of Luciferase Reaction Substrate II to 50 volume of Luciferase Reaction Buffer II. Luciferase Reaction Reagent II can be stored in dark.
3. Prepare 1xCell Lysis Buffer
One volume of 5×Cell Lysis Buffer to 4 volume of ddH2O. 1×Cell Lysis Buffer can be stored at 4oC for 1 month.
4. Prepare Cell Lysate
Wash cells twice with PBS and add 1×Cell Lysis Buffer. Incubate at room temperature for 10 minutes and then scrape the cells. Centrifuge at 12,000 ×g for 10 minutes at 4oC. Transfer the supernatant to a new tube.
5. Assay
Mix 20 μl lysate with 100 μl of Luciferase Reaction Reagent. Place the tube in luminometer to measure the firefly luciferase. Then, add 100 μl of Luciferase Reaction Reagent II to the tube, place the tube in luminometer to measure the renilla luciferase. Record the data.
