Available in lyophilized format.
Substrates provided separately for flexibility.
Detect as little as 0.01 pmol ATP or a single viable cell.
Firefly and Renilla luciferases are widely used as reporter genes for research and drug screening. Our luciferase kits offer excellent sensitivity at a fraction of the cost of other suppliers. Our lyophilized firefly assay kits can be shipped at room temperature to save you even more money. We also offer a wide selection of D-Luciferin and coelenterazine substrates available separately.
Bioluminenscent Enzyme Reactions
Most cells have no intrinsic bioluminescence or luciferase activity, making luciferase reporter assays highly specific and sensitive. The bioluminescent signal of firefly luciferase is generated by the ATP-catalyzed oxidation of D-luciferin into oxyluciferin, which releases light. This is a highly efficient reaction, which makes this system extremely sensitive and linear as a reporter assay. The light emitted by the reaction can be measured by a luminometer, and signal strength can be directly correlated to the amount of firefly luciferase protein expressed.
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
Several commercial luciferase reporter assay kits are available that contain an expression vector containing the luciferase gene. A luciferase reporter assay is usually performed to investigate the strength and regulation of a promoter. This assay has three general steps: 1) clone your promoter of interest upstream of the luciferase gene in an expression vector; 2) transfect cells with the expression vector, and allow cells to grow to express the luciferase; 3) lyse the cells and measure luciferase expression with a luminometer. For this step we recommend our kits described below, which provide lysis reagents, luciferin and buffers optimized for high sensitivity and linearity.
Cell Viability Assays
Bioluminescent assays can be used to monitor cell viability by quantitating ATP. Because ATP is an indicator of metabolically active cells, the number of viable cells can be assessed based on the amount of ATP available.
Reporter Gene Assays
Firefly & Renilla Assay Kits
Our Firefly and Renilla kits offer high sensitivity and linearity comparable to leading competitors. 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 commonly 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. Our assay offers sensitivity and linearity comparable to other two-reporter assays, at less than half the cost. This assay is a flash-type assay, which requires signal to be read in a luminometer immediately after adding reagents.
Features
- 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 for 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. Steady-Luc™ is also available in lyophilized format for economical room temperature shipping and -20°C storage.
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
Cell Viability Assay
ATP-Glo™ Bioluminometric Cell Viability Assay
ATP-Glo™ is a highly sensitive and linear assay that allows quantitation of viable mammalian cells based on their ATP levels. The assay can detect as little as 0.01 pmol ATP or a single viable cell. It is a homogeneous assay for use with cells in culture medium. Note: ATP-Glo™ is a flash-type assay that requires samples to be read in a luminometer immediately after adding the reagent.
Features
- Highly sensitive and linear flash-type assay for cell viability
- Homogeneous assay, add directly to cells in medium
- Add substrate to buffer as needed for consistent performance
- Includes ATP standard
Luciferase Assay Ordering Information & Cross-Reference
| Promega assay | Promega catalog no. | Biotium alternative | Biotium catalog 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 |
ATP-Glo™ Cell Viability Assay | 30020 | Flash | Cell Titer Glo® is a glow type assay (5h half-life); ATP-Glo™ is a flash type assay (signal stable for 1 minute). |
Associated products
Firefly Luciferase Assay Kit 2.0
30085-T, 30085-1, - 30085-2View allHide
Firefly Luciferase Assay Kit (Lyophilized)
30075-1, - 30075-2View allHide
Steady-Luc™ Firefly HTS Assay Kit
30028-1, 30028-2, 30028-3, - 30028-TView allHide
Steady-Luc™ Firefly HTS Assay Kit (Lyophilized)
30028-L1, 30028-L2, - 30028-L3View allHide
Firefly & Renilla Luciferase Single Tube Assay Kit
30081-1, 30081-2, - 30081-TView allHide
Renilla Luciferase Assay Kit 2.0
30082-1, 30082-2, - 30082-TView allHide
ATP-Glo™ Bioluminometric Cell Viability Assay
30020-1, 30020-2, - 30020-TView allHide