The luciferase reporter gene refers to a reporter system that uses luciferin as a substrate to detect the activity of firefly luciferase. Luciferase can catalyze the oxidation of luciferin to oxyluciferin. During the oxidation of luciferin, it will emit bioluminescence.
Firefly luciferase
The most common and common reporter gene is the luciferase of North American firefly photinus pyralis. This protein does not require post-translational modification to obtain enzyme activity. High concentrations (in vivo) are not even toxic, and can be used in prokaryotic and eukaryotic cells.
Amplite™ Luciferase Reporter Gene Detection Kit
Figure 1. In a white 96-well plate with NOVOstar plate reader (BMG Labtech), the Amplite™ luciferase reporter gene detection kit was used to detect luciferase dose response. The kit can detect as low as 0.1pg/well of luciferase within the incubation time of 20min-5h without loss of signal intensity. The half-life exceeds 4 hours.
The Amplite™ Luciferase Reporter Assay Kit (12518) uses a DTTzhuanli-free formulation to quantify the luciferase activity in live cells and cell extracts. The assay is based on firefly luciferase, which is a monomeric 61 kD enzyme that catalyzes the two-step oxidation of luciferin and produces light at 560 nm.
Features of Amplite™ Luciferase Reporter Gene Detection Kit (12518):
With optimized "mixed reading" determination protocol, compatible with HTS liquid handling instruments
With high sensitivity, it can be used for determinations that require low detection limits
A fast, simple and uniform bioluminescence assay for studying gene regulation and function
Compatible with the use of standard cell growth media
Gaussian Luciferase
In recent years, the use of other luciferases (such as Gaussian luciferase) has increased because these reporter genes are small and do not require the presence of ATP. Gaussian luciferase is a 20 kD protein that can catalyze the oxidation of coelenterazine by oxygen to produce light. The bioluminescent enzyme from the marine pod Gaussella can be effectively secreted from mammalian cells after expression.
Amplite™ Gaussian Luciferase Reporter Assay
Figure 2. Comparison of Amplite Gaussian Luciferase Reporter Gene Assay Kit and other brand Gaussian Luciferase Reporter Gene Assay Kit (red line) in 96-well whiteboard using NOVOstar plate reader to measure secreted Gaussian Luciferase Medium Figure.
The Amplite™ Gauss Luciferase Reporter Gene Assay Kit uses a proprietary luminescent formula to quantify the luciferase activity in cell culture media. When the reagent interacts with Gauss luciferase, a luminescent product is produced, which provides strong luminescence.
Features of Amplite Gauss Luciferase Reporter Gene Assay Kit (AAT-12530):
All basic components compatible with HTS liquid handling instruments are provided
They are highly sensitive and can be performed in convenient 96-well and 384-well microtiter plate formats
The "glow type" signal with a half-life of one hour provides a consistent signal among a large number of test panels
Compatible with standard cell growth media
Renilla Luciferase
Renilla luciferase is a 36 kDa protein isolated from Renilla reniformis. Compared with firefly luciferase, Renilla luciferase has different substrate and cofactor requirements. Renilla luciferase uses coelenterazine in the presence of oxygen to produce 480 nm blue light. Similar to firefly luciferase, Renilla luciferase can be used for dual report detection due to differences in substrate requirements and light output.
Amplite™ Renilla Luciferase Reporter Gene Assay
The Amplite™ Renilla Luciferase Reporter Gene Assay Kit provides a fast and sensitive method to detect Renilla luciferase activity in a cell-based assay using a proprietary luminescence formula, as compared to Renilla luciferase After the enzyme interacts, the reagent produces a product with strong light.
Features of Amplite™ Renilla Luciferase Reporter Gene Detection Kit (12535):
The assay is compatible with standard cell growth media
The kit can measure the raw expression or gene expression of wild-type and synthetic hRluc genes
Each kit contains essential components that can facilitate 96-well and 384-well plate testing
Figure 3. The Amplite™ Renilla luciferase reporter gene detection kit was used to measure Renilla luciferase in a white 96-well plate with a NOVOstar microplate reader (BMG Labtech).
Various luciferase substrates, cofactors and physical properties:
