2012 Promega catalogue

Life
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2012
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Cell Signaling
260
Chroma-Glo Luciferase Assay System
Product Size Cat.# Price ($)
Chroma-Glo Luciferase Assay System 10 ml E4910 267.00
For Research Use Only. Not for Use in Diagnostic Procedures.
100 ml E4920 1442.00
Description: The Chroma-Glo Luciferase Assay System and the
Chroma-Luc Vectors provide a method to generate red and green (dual-
color) luminescence from a single sample upon a single-reagent addition.
Filtered measurement of the dual-color luminescence produced by the
Chroma-Luc luciferases permits each reporter to be measured independently
and virtually simultaneously. The Chroma-Glo Assay is in a homogeneous
format that generates luminescence with >30-minute signal half-lives for each
of the Chroma-Luc luciferases, thereby enabling the processing of many
plates without prior sample handling. Use the high-homology Chroma-Luc
luciferases to establish an ideal internal control for normalizing cytotoxicity
in downregulation applications and for decreasing inter- and intrasample
variability. You can also use the reporters to multiplex experimental reporters to
increase the data content from cell-based assays.
Features:
• Measure Dual Reporters Using a Single Substrate Addition: Increase
your accuracy and precision through normalization, or use both reporters to
multiplex experimental measurements. Use filters to spectrally separate the
luminescent signals.
• Establish the Ideal Control or Multiplexed System: Use the highhomology
red and green luciferases to minimize potential RNA and protein
effects on reporter expression.
• Increase Your Throughput: Use the stable luminescence for batch or
continuous processing of multiple plates.
• Perform Fewer Steps: Add Chroma-Luc Reagent directly to cells in
medium, then measure.
• Automate This Assay: Validated automated methods available at:
www.promega.com/automethods/
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store the Chroma-Glo Substrate at –20°C. Store the
Chroma-Glo Assay Buffer below 25°C.
Protocol Part#
Chroma-Glo Luciferase Assay System Technical Manual TM062
Fold Induction
25
20
15
10
5
0
CRE-CBG99luc
NFκB-CBRluc
ISO TNFα ISO +TNFα
Treatment
Using the Chroma-Luc Technology to monitor two independent
experimental signals from the same sample. DNA segments containing
either CRE or the NFκB consensus sequence were cloned into pCBG99-Basic
(Cat.# E1431) or pCBR-Basic (Cat.# E1411). The resulting constructs, pCRE-
CBG99-luc and pNFκB-CBRluc, were cotransfected into 293 cells. At 24
hours post transfection, the cells received one of three treatments: ISO (1μM)/
RO(100μM), TNFα (0.1μg/ml)/RO(100μM), or ISO(1μM)/RO(100μM) plus TNFα
(0.1μg/ml). Only RO(100μM) was added to the Control wells. At six hours post
treatment, cells were harvested and assayed with the Chroma-Glo Reagent.
Relative light units were measured using the Mithras LB940 (Berthold
Technologies) configured with a red filter (610 long pass) and a green filter
(510/60). The red and green signals were deciphered by using the Chroma-
Luc Calculator (available as a downloadable file at:
www.promega.com/chromacalc/). Fold inductions were calculated by
dividing the three treatments by the RO Control.
For complete and up-to-date product information visit: www.promega.com/catalog
4222MA06_3A