2012 Promega catalogue

Cell Signaling
Firefly Luciferase Reporter Systems
ONE-Glo Luciferase Assay System
Product Size Cat.# Price ($)
ONE-Glo Luciferase Assay System 10 ml E6110 249.00
For Laboratory Use.
100 ml E6120 1962.00
1 L E6130 Pls. Enq.
Description: The ONE-Glo Luciferase Assay System provides a highly
sensitive, robust, homogeneous assay for detection of firefly luciferase reporter
gene expression in mammalian cells. Ideally suited for high- and ultrahighthroughput
applications, the ONE-Glo Assay contains a new luciferase
substrate, resulting in a reagent that is more stable, more tolerant to sample
components, and has less odor than standard luciferase assay reagents. These
features ensure that the ONE-Glo Assay provides robust performance and
also eliminates many of the handling inconveniences experienced using other
reporter assays in a high-throughput setting.
Features:
• Simplify Your Assay Optimization: The robust performance, reduced
odor, improved storage, and larger available sizes make assay optimization
even easier and more efficient.
• Store at Room Temperature or at 4°C: The extended stability of the
ONE-Glo Reagent at room temperature and 4°C makes it more
convenient for everyday use.
• Improve Assay Precision: Because the ONE-Glo Reagent is less
sensitive to mixing and dispensing conditions, reproducibility is enhanced.
The assay is ideal for use in high-density (384- and 1536-well)
microplates.
• Get a Brighter, Longer-Lasting Signal: Optimized for batch and
continuous-process handling, the extended bright light output of the ONE-
Glo Assay allows high sensitivity, especially when extended incubation is
required prior to reading results.
• Reduce Unwanted Effects from Sample Components: The novel
chemistry used in the ONE-Glo Assay is less sensitive to culture media,
phenol red, and luciferase inhibitors than other luciferase assays.
Storage Conditions: Store the ONE-Glo Luciferase Assay System
components at –20°C. Please refer to the Technical Manual for other storage
options, including room-temperature storage.
Protocol Part#
ONE-Glo Luciferase Assay System Technical Manual TM292
Luminescence (RLU)
1 × 10 7
1 × 10 6
Bright-Glo Reagent
ONE-Glo Reagent
Steady-Glo ® Reagent
1 × 10
0 20 40 60 80100 120
Time (minutes)
5
ONE-Glo Reagent generates bright and stable luminescence that
can easily be measured for multiple hours. Samples in 96-well plates
consisted of 50μl of purified firefly luciferase (14.9ng/ml with 0.1% Prionex ® )
combined with 50μl of the respective reagent. Luminescence was measured
(1.0 second integration/well) at 3 minutes and periodically for almost 2 hours.
All coefficients of variation were < 3%; n = 3.
For complete and up-to-date product information visit: www.promega.com/catalog
6837MA
Relative Luminescence (%)
100
80
60
40
20
0
ONE-Glo
Reagent
Competitor
(Brite reagent)
ONE-Glo Reagent protects the luciferase reaction in the presence
of resveratrol, a known luciferase inhibitor (Bakhtiarova, A. et al. (2006)
Biochem. Biophys. Res. Comm. 351, 481–4). Luciferase reactions generated
by ONE-Glo Reagent or another bright-type reagent were initiated in the
presence or absence of 10μM resveratrol. Luminescence was initiated and
measured by the method noted in the figure above. The relative luminescence
is the luminescence from reactions containing resveratrol/ luminescence from
reactions without resveratrol x 100; n = 3.
Relative Luminescence (%)
120
100
80
60
40
20
ONE-Glo Reagent
Bright-type
0
0 5 10
Phenol Red (mg/L)
15 20
ONE-Glo Reagent is more tolerant of phenol red than luciferinbased
reagents. Luciferase reactions composed of 14.9ng/ml luciferase
in phenol red-free RPMI medium (with 0.1% Prionex ® ) and ONE-Glo
Reagent or another bright-type reagent were initiated in the presence of
varied amounts of phenol red. Relative luminescence is the luminescence of
reactions containing phenol red/ luminescence from reactions without phenol
red x 100; n = 3.
6841MA
6840MB
261
15
Reporter Assays and Transfection
Section
Contents
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Contents