ONE-Step™ Luciferase Assay System

Catalog #: 60690-1
Size: 10 ml
$115
Catalog #: 60690-2
Size: 100 ml
$795
Catalog #: 60690-5
Size: 500 mL
$3,150
Catalog #: 60690-3
Size: 1 L
$5,995
Background:

Luciferase is the general term given to a class of oxidative enzymes that catalyze reactions that give off light, a process known as bioluminescence. In biology, researchers can take advantage of this reaction and use it as a readout for various biological processes. This has perhaps been exploited most in luciferase reporter cell lines where a promoter region from a gene of interest is placed immediately upstream of the coding sequence for luciferase. In this system, transcriptional activation of the gene of interest leads to a level of luciferase expression that is proportional to the level of gene activation.

One Step Luciferase Assay System


The level of activation is then assessed by lysing the cells and adding the luciferase substrate, D-luciferin. Using the One-Step Luciferase Assay System from BPS Bioscience, this is accomplished in a single step as the lysis buffer has been formulated with D-luciferin. After a brief incubation period, bioluminescence is read on a luminometer.

Description:

The One-Step Luciferase Assay System is designed to be used for high-throughput, sensitive quantitation of firefly luciferase activity in mammalian cell culture. The reagent consists of two components, a Luciferase Reagent Buffer (Component A) and Luciferase Reagent Substrate (Component B). Component A and Component B are combined to form a working solution that contains all the necessary components for cell lysis and luciferase quantitation. This assay system has several features: 

• Sensitive – highly sensitive detection of firefly luciferase activity. 
• Stable – the signal output is stable for more than two hours, providing flexibility with regard to incubation time 
• Convenient – simple one-step, homogeneous protocol. 
• High-throughput – one-step homogeneous protocol minimizes handling steps to support high-throughput screening applications 
• Compatibility – works well with a variety of common media containing 0-10% serum and phenol red. 
• Instrumentation – does not require a luminometer with injectors.

Synonym(s):
Luciferin, Firefly luciferase, luciferase buffer, one step, step, luciferase detection buffer
Format:

Components for 10 ml Size

See datasheet for further information.

Instructions for use:
See product datasheet for detailed protocol.
Storage / Stability:
At least 6 months when stored as directed. Upon first thaw, store in aliquots at -20°C. The reagent may be subjected to several freeze/thaw cycles with no effect on functionality, but it is recommended that freeze/thaw cycles be avoided whenever possible.
Application(s):
• Monitor firefly (Photinus pyralis) luciferase activity in cultured mammalian cells. • High-throughput screening using luciferase reporter cell lines
Notes:

• The reagent has been validated in a 96-well format. Other formats will require scaling and optimization by the end-user. 
• Luciferase Reagent Buffer must be at ~ room temperature (20- 25°C) before use. 
• Luciferase Assay Reagent should be added to cells for at least 5 minutes before measuring luminescence to allow complete cell lysis. 
• For maximal light intensity, measure samples within 1 hour of reagent addition.
• Avoid exposing to excessive heat or light during incubation.
• Different cell lines may exhibit variation in lysis ability and/or luminescence signal and may require slight optimization by the end-user.
• Luminescence signal is affected by assay conditions. Results should be compared between samples measured using the same cell type and media/serum combinations.
• To analyze multiple plates, include a common control sample in each plate and normalize the luminescence of each plate to the control contained in the same plate.
• Background luminescence is a characteristic of luminometer performance, therefore, background luminescence must be subtracted from all readings for accuracy.

Warning(s):
Protect from light. Avoid freeze/thaw cycles.
Scientific Category:
Cell-Based Assay Kits
Product Type:
Reagent
Data shown is lot-specific. Contact us for specific information on other lots.

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$115

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