Description
Luciferase Reporter Gene Assay Kit (BA0181) (BA0181)
The Luciferase Reporter Gene Assay Kit (SKU: BA0181) is a bioluminescent assay for the quantitation of luciferase expression in mammalian, yeast or E. coli cells, using luciferin and ATP as substrates. The reaction produces light which is conveniently measured on a luminometer, and the assay is extremely sensitive, detecting as little as 2 fg of luciferase and as few as 4 cells, with a linear response over seven orders of magnitude. This ultra-sensitive, homogeneous cell-based assay requires only the addition of a single reconstituted reagent to the cells and measurement of light intensity after a short 2-minute incubation. With extended luminescence emission kinetics (half-life 40 minutes), the assay is especially suited to high-throughput screening in 96-, 384- and 1536-well plates. This 1,000-assay format is ideal for gene regulation studies and high-throughput screening of gene modulators. It is intended for research use only.
| Product Name: | Luciferase Reporter Gene Assay Kit (BA0181) |
| SKU: | BA0181 |
| Detection Method: | Bioluminescent (luminometer) |
| Detection Range: | Detection of as little as 2 fg luciferase and as few as 4 cells; linear over seven orders of magnitude |
| Sample Type: | Mammalian, yeast or E. coli cells expressing luciferase |
| Species Reactivity: | All |
| Assay Time: | Within 10 min |
| Kit Size: | 1,000 Assays |
| Equipment Required: | Microplate reader |
| Storage: | Reagent at -20C; Assay Buffer at 2-8C |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
A bioluminescent reporter gene assay for the quantitation of luciferase expression in mammalian, yeast or E. coli cells using luciferin and ATP as substrates. The homogeneous mix-and-measure format combines cell lysis and detection in a single reagent addition, with light measured on a luminometer after a 2-minute incubation. The assay detects as little as 2 fg luciferase and as few as 4 cells, is linear over seven orders of magnitude and has an extended emission half-life of 40 minutes.
- High sensitivity and wide detection range; detects as little as 2 fg luciferase and as few as 4 cells, linear over seven orders of magnitude
- Compatible with routine laboratory and high-throughput formats and all liquid-handling systems
- Fast and convenient homogeneous mix-and-measure assay allowing detection within 10 minutes
- Robust and amenable to HTS with Z-prime factors of 0.6 to 0.8 in 96- and 384-well plates
- Gene regulation studies including gene expression level and promoter characterisation
- Modulation of gene expression by receptors, transcription factors and small molecules
- High-throughput screening for gene modulators in drug discovery
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Plate and culture cells (80 uL) in white opaque 96-well tissue culture plates, including blank control wells with no cells. |
| 2 | Add test compounds and controls (a volume of 20 uL in PBS or culture medium is recommended), mix well and incubate for the desired period (from several hours up to 3 days). |
| 3 | Reconstitute the Reagent by equilibrating the Reagent and Assay Buffer to room temperature, then combining them and mixing by inversion until thoroughly dissolved. |
| 4 | Add 100 uL of reconstituted Reagent (per 80 uL of cell culture) to each well and mix with the cells. Incubate for 2 minutes at room temperature. |
| 5 | Measure luminescence on a luminometer (integration 1-5 sec is typically appropriate). |
The light intensity (RLU) is directly proportional to the luciferase concentration. For dose-response studies, plot the data against compound concentration and determine the EC50 for gene up-regulators or IC50 for gene down-regulators by non-linear regression analysis.
| Component | Quantity | Storage |
| Reagent (solid, in amber tube) | solid | -20C |
| Assay Buffer | 100 mL | 2-8C |
