2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
Section
Contents
Table of
Contents
Cell Signaling
262
ONE-Glo + Tox Luciferase Reporter and
Cell Viability Assay
Product Size Cat.# Price ($)
ONE-Glo + Tox Luciferase Reporter and Cell 1 plate E7110 355.00
Viability Assay
10 plates E7120 2460.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The ONE-Glo + Tox Assay combines luciferase assay chemistry
with a cell viability marker to better understand reporter gene expression in
the context of cell health. The assay uses a two-step, addition-only process to
make these measurements in a single well of a plate, negating the need to run
parallel assays.
The first part of the assay is a nonlytic fluorescence assay (CellTiter-Fluor
Cell Viability Assay) that measures the relative number of live cells in a culture
population after experimental manipulation. The CellTiter-Fluor Assay
measures a conserved and constitutive protease activity within live cells and
therefore serves as a marker of cell viability. The live-cell protease activity
is restricted to intact viable cells and is measured using a fluorogenic, cellpermeant
peptide substrate (glycylphenylalanyl-aminofluorocoumarin; GF-AFC).
The substrate enters intact cells where it is cleaved by the live-cell protease
to generate a fluorescent signal proportional to the number of living cells. This
live-cell protease becomes inactive upon loss of cell membrane integrity and
leakage into the surrounding culture medium. Fluorescence of the free AFC
fluorophore is measured with a microplate reader or CCD imager using an
excitation wavelength of 380–400nm and emission wavelength of 505nm.
The second part of the assay uses the ONE-Glo Luciferase Assay System to
quantify firefly luciferase reporter gene expression from cells made to express
this reporter enzyme. The ONE-Glo Luciferase Assay Buffer and ONE-Glo
Luciferase Assay Substrate, provided with this system, are combined to form
the ONE-Glo Reagent. Ideally suited for high- and ultrahigh-throughput
applications, the ONE-Glo Assay contains a new fluoroluciferin substrate,
resulting in a reagent that is more stable, more tolerant to sample components,
and has less odor than standard luciferase assay reagents.
Luminescence is measured with a microplate reader or CCD imager.
GF–N
H
CF 3
O
Live-cell protease
substrate can cross
the cell membrane.
Active Live-Cell
Protease
Nucleus
O
LIVE-CELL SUBSTRATE:
cell-permeant fluorogenic
substrate for the live-cell
protease (Gly-Phe-AFCoumarin)
H 2N
GF
CF 3
O
Live Cell
The CellTiter-Fluor Cell Viability Assay chemistry portion of the
One-Glo + Tox Assay is a nonlytic fluorescent assay that measures
the relative number of live cells in a culture population after
experimental manipulation.
O
6995MB
Features:
• Efficient: Measure cell viability and firefly luciferase gene expression
within the same assay well. Acquire more data by performing two
measurements on cells in a single assay well.
• Better Biology: Understand reporter gene expression in the context of cell
viability.
• Easy to Perform: The assay uses a simple sequential “add-mix-read”
format.
• Flexible and Automation-Friendly: The volumes of each assay
component can be scaled to meet throughput needs and is amenable to
automation up to 1,536-well format.
Storage Conditions: Store the ONE-Glo + Tox Luciferase Reporter and Cell
Viability Assay components at –20°C. Please refer to the Technical Manual for
other storage options.
Protocol Part#
ONE-Glo + Tox Luciferase Reporter and Cell Viability Assay
Technical Manual
TM356
Steady-Glo ® Luciferase Assay System
Product Size Cat.# Price ($)
Steady-Glo ® Luciferase Assay System 10 ml E2510 187.00
100 ml E2520 1371.00
10 × 100 ml E2550 Pls. Enq.
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: High-throughput quantitation of firefly (Photinus pyralis)
luciferase expression in mammalian cells is commonly performed by batch
processing of 96- and 384-well plates. Steady-Glo ® Luciferase Assay System
is designed for this purpose by providing long-lived luminescence when added
to cultured cells. The homogeneous assay provides signal half-lives of over
5 hours in commonly used cell culture media without prior sample processing.
Throughput rates of several thousand samples per hour may be achieved with
high reproducibility under standard laboratory conditions.
Features:
• Greater Light Output: Results in greater assay sensitivity than other
leading extended-lifetime firefly luciferase assay reagents.
• Improved Assay Precision: Less sensitive to mixing conditions for
improved assay reproducibility in multiwell plates. This is particularly useful
in 384-well plates.
• Convenient: Simply mix buffer with lyophilized substrate and use; there is
no need for thawing or measuring before use.
• Quick: No need to remove culture medium or wash cells prior to adding
assay reagent. Cells may be grown and assayed directly within the same
multiwell plate.
• Easy to Use: Simply add reagent, which contains a cell lysis component,
wait 5 minutes and measure luminescence.
• Robust: Compatible with many tissue culture media, including those
containing up to 10% serum.
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store Steady-Glo ® Luciferase Assay Substrate at –20°C.
Store Steady-Glo ® Luciferase Assay Buffer below 25°C.
Protocol Part#
Steady-Glo ® Luciferase Assay System Technical Manual
TM051
For complete and up-to-date product information visit: www.promega.com/catalog