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Bioluminescent Enzyme Reactions
Most cells have no intrinsic bioluminescence or luciferase activity, making luciferase reporter assays highly specific and sensitive. Firefly luciferase emits light through the ATP-catalyzed oxidation of D-luciferin into oxyluciferin, and signal strength can be directly correlated to the amount of firefly luciferase protein expressed. This is a highly efficient reaction, which makes this system extremely sensitive and linear as a reporter assay.
Renilla luciferase enzyme from the sea pansy catalyzes the oxidation of coelenterazine to generate light. Because firefly and Renilla luciferases use different substrates, they can be used in multiplex assays, typically with one enzyme serving as normalization control for transfection efficiency, and the other enzyme serving as a reporter enzyme.
Reporter Gene Assays
Luciferase reporter systems use an expression vector containing the luciferase gene to investigate the strength and regulation of a promoter in three general steps: 1) clone your promoter of interest upstream of the luciferase gene; 2) transfect cells with the expression vector, and allow cells to grow to express the luciferase; 3) lyse the cells and measure luciferase expression. Our Firefly and Renilla assays have excellent sensitivity and linearity.
Cell Viability Assays
Cell Viability Assay
Steady-ATP™ HTS Cell Viability Assay
The Steady-ATP™ HTS Viability Assay Kit allows highly sensitive quantitation of cellular ATP levels to assess viability with just a single addition of the Steady-ATP™ Assay Mix directly to cells in culture medium. The assay uses an engineered Chemostable Luciferase to oxidize D-Luciferin in the presence of ATP, which emits light. The assay is an improved replacement for the ATP-Glo™ Bioluminometric Cell Viability Assay Kit (Cat. No. 30020).
- Exceptional sensitivity, as low as 16 cells/well in a 384-well plate
- Convenient single-step assay mix
- Compatible for use with high-throughput screening platforms
- Excellent linearity, over 3-4 orders of magnitude
- Stable signal, with a half-life over 5 hours
- Comparable performance to CellTiter-Glo® at a lower cost
Exceptional Sensitivity and Linearity
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Reporter Gene Assays
Firefly & Renilla Assay Kits
Our Firefly and Renilla kits offer high sensitivity and linearity. Unlike other kits that have limited shelf life after reagents are reconstituted, our kits provide substrate separately from reaction buffer, for consistent performance over time. Firefly Luciferase Assay Kit is also available in lyophilized format. These are flash-type assays that require that samples be read in a luminometer immediately after adding reagents.
Features
- High sensitivity and linearity, with low cost
- Add substrates to buffers as needed for consistent performance
- Firefly assay available in lyophilized format
- Water-soluble Aquaphile™ Renilla substrate
Firefly & Renilla Dual Assay Kit
Renilla luciferase is often used as a normalizing transfection control for Firefly reporter assays. Our Firefly & Renilla Single Tube Assay allows you to sequentially measure Firefly and Renilla activities in the same sample in a single tube with sensitivity and linearity comparable to other two-reporter assays. This assay is a flash-type assay that requires signal to be read in a luminometer immediately after adding reagents.
- Assay Firefly & Renilla sequentially in the same sample
- Excellent performance, at a fraction of the cost of other suppliers
- Add substrates to buffers as needed for consistent performance
- Water-soluble Aquaphile™ Renilla substrate
Steady-Luc™ Glow Assay High Throughput Screening
Our Steady-Luc™ Firefly HTS Assay Kit has a signal half-life of about 3 hours, for convenient processing of samples in multi-well plates using a luminescence plate reader. It is a homogeneous assay for use with cells in culture medium.
Features
- Long half-life Firefly assay for multi-well plates
- Homogeneous assay, add directly to cells in medium
- Add substrate to buffer as needed for consistent performance
- Available in lyophilized format
Luciferase Assay Ordering Information & Cross-Reference
| Promega Assay | Promega Cat. No. | Biotium Alternative | Biotium Cat. No. | Assay Type | Notes |
|---|---|---|---|---|---|
| Steady-Glo® Assay System | E2510, E2520, E2550 | Steady-Luc™ Firefly HTS Assay Kit | 30028 | Glow | Signal half-life ~3 hours. Luciferase substrate is added to assay buffer as needed, so signal doesn't drop during kit storage. |
| Steady-Luc™ Firefly HTS Assay Kit (Lyophilized) | 30028L | Glow | Signal half-life ~3 hours. Lyophilized format for convenient shipping and storage. |
||
| Firefly Luciferase Assay System | E1483, E1500, E1501, E4030, E4530, E4550 | Firefly Luciferase Assay Kit 2.0 | 30085 | Flash | Luciferase substrate is added to assay buffer as needed, so signal doesn't drop during kit storage. |
| Firefly Luciferase Assay Kit (Lyophilized) | 30075 | Flash | Lyophilized format for convenient shipping and storage. | ||
| Renilla Luciferase Assay System | E2810, E2820 | Renilla Luciferase Assay Kit 2.0 | 30082 | Flash | Features Biotium's water-soluble Aquaphile™ Coelenterazine. |
| Dual-Luciferase® Assay System (Firefly & Renilla) | E1910, E1960, E1980 | Firefly & Renilla Luciferase Single Tube Assay Kit | 30081 | Flash | Sequentially measure firefly and Renilla luciferase activities in the same sample in one tube. |
| CellTiter-Glo® Luminescent Cell Viability Assay | G7570, G7571, G7572, G7573 | Steady-ATP™ HTS Viability Assay Kit | 30138 | Glow | Signal half-life over 5 hours Convenient single-step assay mix |
