2013 Promega catalogue

Life
Science
Catalog
2013
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Cell Signaling
Firefly Luciferase Reporter Systems
ONE-Glo Luciferase Assay System
Product Size Cat.# Price ($)
ONE-Glo Luciferase Assay System 10 ml E6110 249
For Laboratory Use.
100 ml E6120 1962
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 Assay Optimization: The robust performance, reduced odor,
improved storage, and larger available sizes make assay optimization easy
and efficient.
• Store at Room Temperature or at 4°C: The extended stability of the
ONE-Glo Reagent makes it more convenient for everyday use.
• Improve Assay Precision: The ONE-Glo Reagent is less sensitive to
mixing and dispensing conditions, enhancing reproducibility. 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 allows high
sensitivity, especially for extended incubations.
• Reduce Unwanted Effects from Sample Components: 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
ONE-Glo Luciferase Assay System Technical Manual
Part#
TM292
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 15 20
Phenol Red (mg/L)
6840MB
6841MA
Luminescence (RLU)
1 × 10 7 Bright-Glo Reagent
ONE-Glo Reagent
Steady-Glo ® Reagent
1 × 10 6
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.
1 × 10 5
0 20 40 60 80 100 120
Time (minutes)
6837MA
Section
Contents
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.
Table of
Contents
300
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