2012 Promega catalogue

Cell Signaling
Dual-Glo ® Luciferase Assay System
Product Size Cat.# Price ($)
Dual-Glo ® Luciferase Assay System 10 ml E2920 479.00
For Research Use Only. Not for Use in Diagnostic Procedures.
100 ml E2940 Pls. Enq.
10 × 100 ml E2980 Pls. Enq.
Description: The Dual-Glo ® Luciferase Assay System is a homogeneous
reagent system that enables fast and simple quantitation of a stable
luminescent signal from two reporter genes in a single sample. This convenient
“add-and-read” system generates both firefly and Renilla luciferase luminescence
signals from cells that have not been preconditioned or prelysed. The
Dual-Glo ® Luciferase Assay System provides high Z´-factors for cell-based,
high-throughput screening applications. With the Dual-Glo ® System, internal
controls can be established to minimize sample variability by reducing false
positive and false negative readings caused by nonspecific factors such as
cytotoxicity. In the Dual-Glo ® Luciferase Assay, the activity of the primary
reporter is correlated with the effect of specific stimuli, and the activity of the
co-transfected control reporter provides an internal control to normalize results.
The system is optimized for batch processing both 96- and 384-well plates
and is compatible with a wide variety of mammalian cell culture media.
Features:
• Increased Precision and Accuracy: Normalize primary reporter results
with an internal control, a co-reporter that minimizes effects of cell number
and health, transfection efficiency and nonspecific cellular responses.
• Homogeneous Format: Perform fewer steps. Assay cells directly in
growth medium for both reporters. No centrifugation or lysis step is
required.
• Stable Signal: Obtain flexibility for either batch or continuous processing
of 96- and 384-well plates. Each luminescent signal may be measured for
up to 2 hours after reagent addition.
• Convenience: Screen efficiently with this simple, two-step assay that is
ideal for any luminometer. On-board injectors not required.
• Wide Dynamic Range: Analyze high and low reporter activity without
sample dilution. Linear over at least 6 logs of enzyme concentration for
each reporter.
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store Dual-Glo ® Substrates at –20°C. Store Dual-Glo ®
Buffers below 25°C.
Protocol Part#
Dual-Glo ® Luciferase Assay System Technical Manual
TM058
Dual-Luciferase ® Reporter Assay System
Product Size Cat.# Price ($)
Dual-Luciferase ® Reporter Assay System
Dual-Luciferase
100 assays E1910 398.00
® Reporter Assay System
10-Pack
Dual-Luciferase
1,000 assays E1960 2555.00
® Reporter 1000 Assay
System
Available Separately
1,000 assays E1980 2239.00
Passive Lysis 5X Buffer 30 ml E1941 134.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The Dual-Luciferase ® Reporter (DLR) Assay System provides
an efficient means of performing two reporter assays. In the DLR Assay, the
activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis or sea pansy)
luciferases are measured sequentially from a single sample. The firefly
luciferase reporter is measured first by adding Luciferase Assay Reagent II (LAR
II) to generate a luminescent signal lasting at least one minute. After quantifying
the firefly luminescence, this reaction is quenched, and the Renilla luciferase
reaction is initiated simultaneously by adding Stop & Glo ® Reagent to the same
sample. Both assays can be completed in about 4 seconds using a
For complete and up-to-date product information visit: www.promega.com/catalog
luminometer with reagent auto-injectors. In the DLR Assay System, both
reporters yield linear assays with attomole (100μl/well), additional Passive Lysis Buffer may be purchased separately.
Features:
• Greater Accuracy: Renilla luciferase internal control allows for more
accurate results.
• Convenient: Samples don’t have to be split, which saves plates and time.
• Sensitive: Allows study of weak promoters, low-level expression/regulation,
and expression in cells that transfect poorly.
• Linear: Range extends 7 logs; very active samples typically do not need
dilution.
Storage Conditions: Store at –20°C.
Protocol Part#
Dual-Luciferase ® Reporter Assay System Technical Manual TM040
Dual-Luciferase ® Reporter 1000 Assay System Technical Manual TM046
Percent Optimal Transfection
120
80
40
0
Optimal Transfection
Suboptimal Transfection
None Protein Vector
Method of Normalization
Effect of transfection conditions on reporter results analyzed using
different normalization methods. HEK 293 cells were transfected with
pGL4.13[luc2/SV40] expressing firefly luciferase and pGL4.74[hRluc/TK]
expressing Renilla luciferase. Transfections were performed using both
optimal and suboptimal lipid:DNA ratios (indicated as Optimal and Suboptimal
Transfection conditions). Firefly and Renilla luciferase activities were measured
using the Dual-Luciferase ® Reporter Assay System (Cat.# E1960). Protein
concentrations were determined using the Coomassie ® Plus Bradford Reagent
(Pierce). Firefly luciferase data were either not normalized (None), normalized
to total protein (Protein), or normalized to Renilla luciferase activity (Vector).
Data represent the average ± standard deviation of triplicate samples and
are expressed as a percent of the optimal transfection for each normalization
condition.
6398MA
259
15
Reporter Assays and Transfection
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