The firefly luciferase assay is commonly used as a tool to study gene regulation at transcriptional level. This approach has been widely used because it is sensitive, convenient and able for quantitative measurements. Beyotime provides a variety of high-quality firefly luciferase assay systems.
Based on those products, we have now developed the Bio-Lumi™ Firefly Luciferase Assay System, abbreviated as Bio-Lumi. This product uses a new firefly luciferase substrate which is more tolerant of sample components than standard assay reagents, enabling a more stable reaction and the reproducibility of experimental results. Therefore, the Bio-Lumi is ideal for ADCC Reporter Bioassay and other firefly luciferase reporter assays.
The Bio-Lumi assay system is provided in two forms: the ready-to-use Bio-Lumi™ Firefly Luciferase Assay System (RG042) and the lyophilized Bio-Lumi™ II Firefly Luciferase Assay System (RG043). Their performance in analyzing the activity of firefly luciferase is exactly the same. RG042 must be stored at -80℃. Its detection performance gradually decreases when stored at -20℃. RG043 is a lyophilized version of RG042. The lyophilized substrate is very stable at -20℃, but must be dissolved in Assay Buffer prior to use.
Features:
Stable Luminescence with High Intensity: The luminescence signal fluctuates no more than 5% within 10 minutes, no more than 15% within 30 minutes, and no more than 30% within 60 minutes.
Simple Operation and Rapid Detection: The assay reagent can be added directly to cell culture at an equal volume without the need of cell washing or medium changing. The measurement of a 96-well plate can be completed in less than 2 minutes using a compatible microplate reader.
High Flexibility: Suitable for a small number of tests and high-throughput screening. Both cell culture and cell lysate can be used for this assay system.
High Compatibility: Compatible with various common culture media. Phenol red, fetal calf serum or calf serum less than 10%, DMSO or ethanol less than 2% in normal culture media have basically no effect on the assay. Commonly used salts or metal ions at normal concentrations do not affect the assay either.
Highly Sensitivity and Wide Linear Range: When a 96-well plate is used, this assay usually gives good linearity between luminescence and cell numbers less than 200,000 (Figure 1). The maximum number of cells in the linear range may vary for different cells or different expression levels of luciferase.
Figure 1. Luminescent signal detection of different numbers of HeLa cells transfected with plasmids carrying the firefly luciferase reporter gene by the Bio-Lumi™ Firefly Luciferase Assay System. Transfection efficiency is about 60-70% as shown with the co-transfected EGFP-expression plasmids. Actual readings may vary depending on the cell type, transfection efficiency, expression plasmid or different instrument used.
Typical Figures:
Bio-Lumi has comparable performance in both detection sensitivity and luminescence stability with the well received assay system from Competitor P. Its detection sensitivity is even better than Competitor P, with the luminescence intensity about 30% higher than that of Competitor P (Figure 3A). The luminescent signal stability is basically the same as that of Competitor P (Figure 3B).
Figure 2. Firefly luciferase assay of HeLa cells transfected with firefly luciferase expression plasmids by the Bio-Lumi (Beyotime, #RG042) and the Competitor P. A. The chemiluminescence intensity generated by Bio-Lumi and Competitor P; B. The stability of chemiluminescence generated by Bio-Lumi and Competitor P. Actual readings may vary depending on the cell type, transfection efficiency, expression plasmid or different instrument used. Values in the graphs are for reference only.
Selection Guide:
Please refer to the table below for the comparison of five types of Beyotime Firefly Luciferase Assay System.
Use Bright-Lumi if a strong luminescent signal is desired and the signal can be measured within 30 minutes.
Use Steady-Lumi when testing a large amount of samples and signal stability is required.
Use One-Lumi if both high signal intensity and signal stability are needed.
Use Bio-Lumi for ADCC Reporter Bioassay requiring both high signal intensity and stability of reactions.
| Catalog No. | RG042 | RG043 | RG052 | RG056 | RG059 |
| Product Abbreviation | Bio-Lumi | Bio-Lumi II | Bright-Lumi II | One-Lumi II | Steady-Lumi II |
| Convenience | Very Convenient | Convenient | Convenient | Convenient | Convenient |
| Sensitivity | Very Sensitive | Very Sensitive | Exceptionally Sensitive | Very Sensitive | Sensitive |
| Signal Intensity | Highest | Highest | Highest | Higher | High |
| Signal Half-life Time | 1-2 hours | 1-2 hours | 0.5-1 hour | 1-2 hours | 5-6 hours |
| Product Stability | Very Stable | Very Stable | Very Stable | Very Stable | Very Stable |
| Linear Range (number of cells) | 50 - 100,000 | 50 - 100,000 | 50 - 100,000 | 50 - 100,000 | 50 - 200,000 |
| Recommend index | ★★★★★ | ★★★★★ | ★★★★★ | ★★★★ | ★★★★ |
Product Information：
| Catalog No. | Product Name | Size | Price |
| RG042S | Bio-Lumi™ Firefly Luciferase Assay System | 100 T | |
| RG042M | Bio-Lumi™ Firefly Luciferase Assay System | 1000 T | |
| RG043S | Bio-Lumi™ II Firefly Luciferase Assay System | 100 T | |
| RG043M | Bio-Lumi™ II Firefly Luciferase Assay System | 1000 T |