Thermo Scientific™ Pierce™ Firefly Luc One-Step Glow Assay Kit

Perform firefly luciferase assays in high-throughput screening applications.

The Thermo Scientific Pierce Firefly One-Step Luc Glow Assay Kit measures firefly luciferase activity in mammalian cell culture using a single reagent-addition step, making it ideal for high-throughput screening (HTS) applications.

This glow-type luciferase assay kit is designed for use with cultured mammalian cells that are transfected with vectors expressing the appropriate firefly luciferase reporter enzyme. Several features make the kit especially suitable for luminometers without injectors and for high-throughput screening (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 wells containing the transfected, treated cells. The stable signal output (glow) of this system provides flexibility with regard to incubation time. When used with Thermo Scientific Firefly Luc Plasmids, the kit provides a highly sensitive bioluminescent reporter assay system for the detection of promoter or pathway activity.

Highlights:

• Sensitive - highly sensitive detection of firefly luciferase activity
• Convenient - one-step, mix-and-read protocol; no additional reagents required
• Stable - increased signal half-life and stability compared to flash assays
• Simple - does not require a luminometer with injectors
• Automation-friendly - amenable to high throughput assays

Includes:
Assay buffer and substrate solution (100X)
Requires:
Firefly luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers.
Applications:

• Promoter studies for analyzing cis- and trans-acting regulatory elements
• Drug screening
• siRNA and miRNA screening
• Multiplexed assays to study off-target effects
• Protein localization reporter assays
• Signal transduction pathway analysis

Product Details:
Firefly luciferase is a 62kDa protein produced in nature by several species of the Lampyridae family of beetles, which includes the genera Photinus and Luciola. Thermo Scientific Red Firefly Luciferase has an emission range of 560-700nm (λmax = 613nm). Bioluminescent signal from firefly luciferase originates from the oxidation of D-Luciferin. Light output captured using a luminometer can be correlated with the amount of firefly luciferase protein produced and used to determine the activity of the promoter driving firefly luciferase expression. Firefly luciferase bioluminescence provides researchers with a non-radioactive assay for measuring transcriptional activity of regulatory elements. Firefly luciferase reporter activity can be used to determine promoter activity, to study upstream pathway activity in the cell, or to determine effect of activators or inhibitors.

Type

Assay Kit

Target

Luciferase, Firefly Luciferase, Cypridina Luciferase

Sufficient For

1000 Luciferase Assays in Microplate Wells

Quantity

1000 Reaction Kit

Content And Storage

• Firefly Luc One-Step Substrate, Lyophilized (makes 1 mL of 1X), store at -20°C
• Firefly Luc One-Step Assay Buffer, 115 mL, store at -20°C

Upon receipt store kit at -20°C.

Assay

Reporter Enzyme, Luciferase Reporter Assay

Compatible Cells

Mammalian Cells

Detection Method

Bioluminescence

For Use With (Equipment)

Luminometer (Microplate)

Format

384-well plate, 96-well plate

Label Type

Enzyme Labeled

Substrate Properties

Chemical Substrate

Technique

Enhanced Chemiluminescence

Substrate Type

Luciferase Substrate

Substrate

D-Luciferin

For Research Use Only. Not for use in diagnostic procedures.