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The Thermo Scientific™ Pierce™ Luciferase reporter genes are human codon optimized and offered in a plasmid vector that is optimized for mammalian expression. See the table below for their key features:
| Cypridina Luciferase | Gaussia Luciferase | Gaussia-Dura Luciferase | Green Renilla Luciferase | Red Firefly Luciferase | TurboLuc™
|
Substrate | Vargulin | Coelenterazine | Coelenterazine | Coelenterazine | D-luciferin | Coelenterazine |
Sample type | Media/Lysate | Media/Lysate | Media/Lysate | Lysate | Lysate | Lysate |
Relative brightness in flash assay | 107 (media)/ 107 (lysate) | 107 (media)/ 107 (lysate) | 107 (media)/ 107 (lysate) | 107 | 105 | NA |
Average half-life in flash assay | 30 min | <5 min | NA | 5 min | >30 min | NA |
Average half-life in glow assay | 1 hr | NA | 1 hr | 3 hr | 2 hr | 2 hr |
We offer the following luciferase assays (each available in Glow and Flash formats) that provide a highly sensitive assay for transcriptional activity of regulatory elements in mammalian cell culture media and whole cell lysate:
See the table below for the Luciferase Glow assay and Luciferase Flash assay kit options we offer:
Luciferase Assay Kit | Reaction size | Cat. No. |
Thermo Scientific™ Pierce™ Gaussia Luciferase Flash Assay Kit | 100 reactions | 16158 |
Thermo Scientific™ Pierce™ Gaussia Luciferase Flash Assay Kit | 1000 reactions | 16159 |
Thermo Scientific™ Pierce™ Gaussia Luciferase Glow Assay Kit | 100 reactions | 16160 |
Thermo Scientific™ Pierce™ Gaussia Luciferase Glow Assay Kit | 1000 reactions | 16161 |
Thermo Scientific™ Pierce™ Renilla Luciferase Flash Assay Kit | 100 reactions | 16164 |
Thermo Scientific™ Pierce™ Renilla Luciferase Flash Assay Kit | 1000 reactions | 16165 |
Thermo Scientific™ Pierce™ Renilla Luciferase Glow Assay Kit | 100 reactions | 16166 |
Thermo Scientific™ Pierce™ Renilla Luciferase Glow Assay Kit | 1000 reactions | 16167 |
Thermo Scientific™ Pierce™ Cypridina Luciferase Flash Assay Kit | 100 reactions | 16168 |
Thermo Scientific™ Pierce™ Cypridina Luciferase Flash Assay Kit | 1000 reactions | 16169 |
Thermo Scientific™ Pierce™ Cypridina Luciferase Glow Assay Kit | 100 reactions | 16170 |
Thermo Scientific™ Pierce™ Cypridina Luciferase Glow Assay Kit | 1000 reactions | 16171 |
Thermo Scientific™ Pierce™ Firefly Luciferase Flash Assay Kit | 100 reactions | 16174 |
Thermo Scientific™ Pierce™ Firefly Luciferase Flash Assay Kit | 1000 reactions | 16175 |
Thermo Scientific™ Pierce™ Firefly Luciferase Glow Assay Kit | 100 reactions | 16176 |
Thermo Scientific™ Pierce™ Firefly Luciferase Glow Assay Kit | 1000 reactions | 16177 |
Gaussia and Cypridina Luciferase assayscan be tested on both media and cell lysate sample types. Renilla and Firefly Luciferase assays can only be used on cell lysate as a sample type.
We always recommend the luciferases to be read without filters for single luciferase assays. Use of filters reduces the amount of signal captured that may lead to decrease in sensitivity. Filters are required only for dual-spectral assays.
The earliest detectable signal will depend on the strength of the promoter. In general, with a strong promoter (such as CMV), significant signal over background can be seen in as little as 20 minutes. However, with a weaker inducible promoter, significant signal over background may take 1-2 hours after induction.
Gaussia, Gaussia-Dura, and Cypridina Luciferase are stable in the culture media for greater than 24 hours. Generally, after transfection of CMV driven constructs, we see ever increasing secreted signal between 24 and 72 hours.
The below table provides a guideline based on sample type and luciferase of choice. A luminometer with injectors is recommended for Flash assays.
Single assay | Isolate Luc from | Substrate | Assay type | Recommended filters |
Cypridina | Media or lysate | Vargulin | Flash, Glow |
No filter needed. Collect total light output. |
Gaussia | Media or lysate | Coelenterazine | Flash | |
Gaussia-Dura | Media or lysate | Coelenterazine | Glow | |
Green Renilla | Lysate | Coelenterazine | Flash, Glow | |
Red Firefly | Lysate | D-Luciferin | Flash, Glow |
The three luciferase reporters, Gaussia, Cypridina and Renilla Luciferase are paired with Firefly Luciferase as a normalization control. For example, Gaussia Luciferase acts as an experimental reporter with constitutively active Red Firefly Luciferase as a normalization control. This reporter-and-control combination enables simultaneous monitoring of experimental reporter and control luciferase activities in a single-read assay without the need for two-step addition of substrate reagents or quenching. The assay working solution contains substrates for both luciferases, and the reactions occur simultaneously with flash-type kinetics. The resulting luminescent signals are spectrally resolvable using filters. In a single sample, researchers can assay transcriptional activity of regulatory elements, signal transduction pathways, and effects of activators or inhibitors.
Luciferease Dual Assay Kit
| Reaction Size | Cat. No. |
Pierce™ Gaussia-FireflyLuciferase Dual Assay Kit
| 100 reactions | 16181 |
Pierce™ Gaussia-FireflyLuciferase Dual Assay Kit
| 1000 reactions | 16182 |
Pierce™ Cyprdina-FireflyLuciferase Dual Assay Kit
| 100 reactions | 16183 |
Pierce™ Cyprdina-FireflyLuciferase Dual Assay Kit
| 1000 reactions | 16184 |
Pierce™ Renilla-FireflyLuciferase Dual Assay Kit
| 100 reactions | 16185 |
Pierce™ Renilla-FireflyLuciferase Dual Assay Kit
| 1000 reactions | 16186 |
No, our dual spectral Luciferase assay kits are not useable with Promega’s Firefly/Renilla reporter vectors. This is because our Firefly Luciferase emits in red range and our Renilla Luciferase emits in green range, while Promega’s Firefly Luciferase emits in green range and their Renilla Luciferase emits in blue range.
Promega offers a number of luciferase assays, but due to Promega’s licenses for their products, we cannot recommend the use of any of our reagents with their vectors or our vectors with their reagents.
TurboLuc ™ Luciferase One-Step Glow Assay Kit measures activity in mammalian cells with a single reagent-addition step, making it ideal for high-throughput screening (HTS) applications. Several features of the TurboLuc ™ Luciferase One-Step Glow Assay Kit make it suitable for luminometers without injectors and for HTS applications. The convenient, one-step, homogenous protocol minimizes handling steps to support the use of automation. Simply combine the supplied substrate solution and assay buffer to make a single working solution, and then add it to microplate containing the transfected, treated cells. The stable signal output (glow enzyme kinetics) of this system provides flexibility with regard to lead time before read. When used with Thermo Scientific™ TurboLuc™16 Luciferase vectors, the TurboLuc ™ Luciferase One-Step Glow Assay Kit provides a highly sensitive bioluminescent reporter assay system for the detection of promoter or pathway activity.
There are three vectors available for each expressed luciferase; pMCS, pCMV, and pTK. We recommend using pMCS to measure the activity of the reporter, and pCMV, and pTK as positive control vectors. For high level constitutive expression of Luciferase, we recommend using the CMV (Cytomegalovirus) promoter vector and for low level constitutive expression of luciferase, we recommend using the pTK (thymidine kinase) promoter vector.
See the table below for the various luciferase vector options we offer:
Name of vector | Size | Cat. No. |
pMCS-Gaussai Luc | 10 µg | 16146 |
pCMV-Gaussia Luc | 10 µg | 16147 |
pTK-Gaussia Luc | 10 µg | 16148 |
pMCS-Cypridina Luc | 10 µg | 16149 |
pCMV-Cyrpidina Luc | 10 µg | 16150 |
pTK-Cypridina Luc | 10 µg | 16151 |
pMCS-Green Renilla Luc | 10 µg | 16152 |
pCMV-Green Renilla Luc | 10 µg | 16153 |
pTK-Green Renilla Luc | 10 µg | 16154 |
pMCS-Firefly Luc | 10 µg | 16155 |
pCMV Firefly Luc | 10 µg | 16156 |
pTK Firefly Luc | 10 µg | 16157 |
pMCS Gaussia-Dura Luc | 10 µg | 16190 |
pCMV Gaussia-Dura Luc | 10 µg | 16191 |
pTK Gaussia-Dura Luc | 10 µg | 16192 |
We offer three vectors with commonly used promoters built into the vectors:
Vector-specific information can be found in the lot-specific Certificate of Analysis on the website
Thermo Scientific™ M-PER Mammalian Protein Extraction Reagent (Cat. No.78503) may be used to prepare lysate from washed adherent or pelleted cells, and then a portion of the lysate can be used to assay for beta-galactosidase activity.
For Research Use Only. Not for use in diagnostic procedures.