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Luciferases

The most versatile and common reporter gene is the luciferase of the North American firefly photinus pyralis. The protein requires no posttranslational modification for enzyme activity. It is not toxic at high concentrations (in vivo) and can be used in prokaryotic or eukaryotic cells.

 

 

Amplite® Luciferase Reporter Gene Assay



Luciferase dose-response was measured with Amplite® Luciferase Reporter Gene Assay Kit in a white 96-well plate with a NOVOstar plate reader (BMG Labtech). The kit can detect as low as 0.1pg/well luciferase with 20 minutes to 5 hours incubation without losing signal intensity. The integration time was 1 second. The half-life is more than 4 hours.
Amplite® Luciferase Reporter Gene Assay Kits use a proprietary DTT-free formulation to quantify luciferase activity in live cells and cell extracts. The assay is based on firefly luciferase, a monomeric 61 kD enzyme that catalyzes two-step oxidation of luciferin, yielding light at 560 nm. The formulation provided in this kit generates a luminescent product that gives strong luminescence upon interaction with luciferase. Kits include all the essential components with an optimized "mix-and-read" assay protocol compatible with HTS liquid handling instruments. They have high sensitivity and can be used for assays that require a low detection limit. The kits have a fast, simple, and homogeneous bioluminescence assay for studying gene regulation and function. The assay is compatible with the use of standard cell growth media.







Table 1. Product ordering information for Amplite® Luciferase Reporter Gene Assay kits.
Product Unit Size Cat No.
Amplite® Luciferase Reporter Gene Assay Kit *Bright Glow* 1 plate 12518
Amplite® Luciferase Reporter Gene Assay Kit *Bright Glow* 10 plates 12519
Amplite® Luciferase Reporter Gene Assay Kit *Bright Glow* 100 plates 12520

 

Guassia Luciferase



Sectreated Gaussia Luciferase culture medium was measured with Amplite® Gaussia Luciferase Reporter Gene Assay Kit (blue line) and a commercially available Gaussia Luciferase Assay Kit (red line) in a white 96-well plate with a NOVOstar plate reader (BMG Labtech).
In recent years, there has been an increase in the use of other luciferases, such as Gaussia luciferase, since these reporters are smaller and do not require the presence of ATP. Gaussia luciferase, which is a 20 kD protein, catalyzes coelenterazine oxidation by oxygen to produce light. The bioluminescent enzyme derived from the marine copepod Gaussia princeps is efficiently secreted from mammalian cells upon expression.

Amplite® Gaussia Luciferase Reporter Gene Assay


Amplite® Gaussia Luciferase Reporter Gene Assay Kits use a proprietary luminogenic formulation to quantify luciferase activity in cell medium. The formulation generates a luminescent product that gives strong luminescence upon interaction with Gaussia luciferase. The kits provide all the essential components that are compatible with HTS liquid handling instruments. They have high sensitivity 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 across many assay plates. The assay is compatible with standard cell growth media.

Table 2. Product ordering information for Amplite® Gaussia Luciferase Reporter Gene Assay kits.
Product Unit Size Cat No.
Amplite® Gaussia Luciferase Reporter Gene Assay Kit *Bright Glow* 1 plate 12530
Amplite® Gaussia Luciferase Reporter Gene Assay Kit *Bright Glow* 10 plates 12531
Amplite® Gaussia Luciferase Reporter Gene Assay Kit *Bright Glow* 100 plates 12532

 

Renilla Luciferase



Renilla Luciferase was measured with Amplite® Renilla Luciferase Reporter Gene Assay Kit in a white 96-well plate with a NOVOstar plate reader (BMG Labtech).
Renilla luciferase is a 36 kDa protein isolated from the sea pansy (Renilla reniformis). Compared to firefly luciferase, Renilla luciferase enzymes differ in their substrate and cofactor requirements. Renilla luciferase uses coelenterazine in the presence of oxygen to produce a blue light of 480 nm. Like firefly luciferase, Renilla luciferase can be used in dual-reporter assays due to their differences in substrate requirements and light output.

Amplite® Renilla Luciferase Reporter Gene Assay


Amplite® Renilla Luciferase Reporter Gene Assay Kits offer a fast and sensitive method to detect Renilla luciferase activity in cell-based assays using a proprietary luminogenic formulation. Upon interaction with Renilla luciferase, this formulation generates a product with strong luminescence. The "glow-type" signal with a half-life of one hour provides a consistent signal across many assay plates. The assay is compatible with standard cell growth media. The kits enable the measurement of primary expression or gene expression with wild-type and synthetic hRluc genes. Each kit includes all the essential components and can be performed in a convenient 96-well and 384-well microtiter-plate format.

Table 3. Product ordering information for Amplite® Renilla Luciferase Reporter Gene Assay kits.
Product Unit Size Cat No.
Amplite® Renilla Luciferase Reporter Gene Assay Kit *Bright Glow* 1 plate 12535
Amplite® Renilla Luciferase Reporter Gene Assay Kit *Bright Glow* 10 plates 12536
Amplite® Renilla Luciferase Reporter Gene Assay Kit *Bright Glow* 100 plates 12537

Table 4. Luciferase specificity, cofactor requirements, and physical characteristics.
Organism Luciferase Size (kDa) Substrate Requires Secreted
Photinus pyralis North American firefly luciferase 61 D-luciferin Mg, ATP No
Luciola cruciata Japanese firefly (Genji-botaru ) luciferase 64 D-luciferin Mg, ATP No
Luciola italica Italian firefly Luciferase 64 D-luciferin Mg, ATP No
Luciola lateralis Japanese firefly (Heike) luciferase 64 D-luciferin Mg, ATP No
Luciola mingrelica East European firefly luciferase 64 D-luciferin Mg, ATP No
Photuris pennsylvanica Pennsylvania firefly luciferase 64 D-luciferin Mg, ATP No
Pyrophorus plagiophthalamus Click beetle luciferase 64 D-luciferin Mg, ATP No
Phrixothrix hirtus Railroad worm luciferase 64 D-luciferin Mg, ATP No
Renilla reniformis Renilla luciferase 36 Coelenterazine N/A No
Rluc8 (mutant of Renillaluciferase) 36 Coelenterazine N/A No
Green Renilla luciferase 36 Coelenterazine N/A No
Gaussia princeps Gaussia luciferase 20 Coelenterazine N/A Yes
Gaussia-Dura luciferase 20 Coelenterazine N/A Yes
Cypridina noctiluca Cypridina luciferase 62 Vargulin/Cypridinaluciferin N/A Yes
Cypridina hilgendorfii Cypridina (Vargula) luciferase 62 Vargulin/Cypridinaluciferin N/A Yes
Metridia longa Metridia luciferase 23.8 Coelenterazine N/A Yes
Oplophorus gracilorostris OLuc 19 Coelenterazine N/A Yes