Super sensitive flash-type or long half-life glow assays.
Measure Firefly & Renilla in the same tube.
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.
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 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.
Several commercially 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.
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.
Bioluminescent reactions catalyzed by firefly luciferase (A) and Renilla luciferase (B).
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.
Comparison of linear ranges of Biotium’s Firefly & Renilla Single Tube Assay and Promega’s Dual Luciferase® Reporter Assay System (DLR™). Luciferase reactions were performed according to manufacturers instructions using titration curves of purified recombinant QuantiLum® recombinant firefly luciferase (Promega) or purified recombinant Renilla luciferase (Ray Biotech). Luciferase enzymes were diluted in 1X passive lysis buffer from the assay kit tested. Background from reagents without enzyme added was subtracted from luminescence values. Both assay systems showed comparable sensitivity and dynamic range.
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.
Firefly & Renilla dual assay format.
Example of Firefly & Renilla Luciferase Single Tube Assay using HeLa cells cells transfected with firefly luciferase alone (firefly only), firefly + Renilla, or no transfection (no tfx). Quenched firefly is the residual firefly signal after adding Renilla working solution to firefly-only transfected cells.
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 homogenous 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.
Luminescence signal over time in firefly luciferase transfected CHO-K1 cells in the Steady-Luc™ Lyophilized 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 homogenous 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.
Quantitation of an ATP standard curve (red squares), or a dilution series of live Jurkat cells (blue diamonds) with the ATP-Glo™ Assay Kit.
Luciferase substrate is added to assay buffer as needed, so signal doesn’t drop during kit storage.
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.
Renilla Luciferase Assay System E2810, E2820 Renilla Luciferase Assay Kit 2.0 30082 Flash Features Biotium’s water-soluble Aquaphile™ Coelenterazine.