What are the advantages of using TurboFect Transfection Reagent (Cat. No. R0533) over other transfection reagents with the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186)?
The Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) luciferase vectors were optimized with TurboFect. However, it should be feasible to replace TurboFect with another transfection reagent if your cells tolerate the transfection reagent better and give you acceptable transfection rates.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can I store my Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) cell lysates at -20 degrees C to test later?
The stability of the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) may be inconsistent from sample to sample at -20 degrees C. We recommend snap-freezing lysates at -80 degrees C with protease inhibitors to help prevent protein degradation.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can I use the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) with a luminometer that is not equipped with reagent injectors?
No. All Luciferase Dual Assay Kits, such as the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186), are flash-based assays. Flash luminescence reactions give a very bright signal over a short reaction time. Luciferase assays that are flash-based require the use of a luminometer or other luminescence-monitoring instrument equipped with automated reagent injectors.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
I got a high background signal when I used a Pierce Gaussia/Cypridina/Renilla -Firefly Dual Assay Kit. Why is this?
Here are possible causes and solutions:
- Non-specific oxidation of substrate: Use a new sample; Avoid repeated freezing and thawing of the sample.
- Control sample is contaminated: Change pipette tips after each well; Reduce shaker speed during the cell lysis step to avoid contaminating the wells.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
I got a high signal when I used a Pierce Gaussia/Cypridina/Renilla -Firefly Dual Assay Kit. Why is this?
This could be due to high luciferase expression. Here are some suggestions:
Reduce incubation time before collecting samples. Decrease the integration time on the instrument.Dilute the sample:
Note: A low sample volume can increase assay variability. Dilute the sample and use the recommended volume of 10-20 µL per assay.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.