2012 Promega catalogue

Cell Signaling
MultiTox-Fluor Multiplex Cytotoxicity Assay
Product Size Cat.# Price ($)
MultiTox-Fluor Multiplex Cytotoxicity Assay 10 ml G9200 322.00
For Laboratory Use.
5 × 10 ml G9201 1195.00
2 × 50 ml G9202 1791.00
Description: The MultiTox-Fluor Multiplex Cytotoxicity Assay is a singlereagent-addition,
homogeneous, fluorescent assay that measures the number
of live and dead cells simultaneously in culture wells. The assay simultaneously
measures cell viability and cytotoxicity by detecting two distinct protease activities.
The live-cell protease activity is restricted to intact viable cells and is measured
using a fluorogenic, cell-permeant peptide substrate (GF-AFC Substrate).
The substrate enters intact cells where it is cleaved to generate a fluorescent
signal proportional to the number of living cells. This live-cell protease activity
marker becomes inactive upon loss of membrane integrity and leakage into the
surrounding culture medium. A second, cell-impermeant, fluorogenic peptide
substrate (bis-AAF-R110 Substrate) is used to measure dead-cell protease
activity that has been released from cells that have lost membrane integrity.
Features:
• Measure the Number of Live and Dead Cells in Culture: “Add-mixmeasure”
protocol eliminates parallel plate processing and reduces cell
culture costs.
• Normalize Data for Cell Number: The ratio of live:dead cells is independent
of cell number and normalizes data, making results more comparable
well-to-well, plate-to-plate, day-to-day.
• Reduce False-Positive and -Negative Results: Complementary liveand
dead-cell measures with independent chemistries serve as internal
controls for each other.
• Get More Data from Every Well: Multiplex the MultiTox-Fluor Assay with
most Promega bioluminescent cell-based apoptosis or genetic reporter
assays.
• Reduce Assay Variability: The homogeneous “add-mix-measure”
protocol avoids the cumulative error associated with multistep protocols.
Storage Conditions: Store at –20°C.
Protocol Part#
MultiTox-Fluor Multiplex Cytotoxicity Assay TB348
For complete and up-to-date product information visit: www.promega.com/catalog
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MultiTox Assay Protocol
Active Live-Cell
1) Treat cells with the e
potential cytotoxic totoxic c agent. aagent.
2) Add substrates. tes.
GF– LIVE-CELL ELL SUBSTRATE:
STRATE:
cell permeant meant fluorogenic fluoro luorogeni enic
substrate for f the h li live-cell ll
protease (Gly-Phe-AFCoumarin)
AAF–
Active Live-Cell
Live-cell protease
substrate can cross
Nucleus the cell membrane;
dead cell protease
substrate cannot.
Live Cell
3) Measure fluorescence/luminescence.
Live
Cells
Protease
Dead
Cells
Active Dead-Cell
Protease
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Live
Cells
Protease
Dead
Cells
Protease
Active Dead-Cell
DEAD-CELL DEAD CELL SUBSTRATE:
SUBSTRATE:
cell cell impermeant im impermeant ermeant substrate
for f the h d ddead-cell d ll protease
(Ala-Ala-Phe-rhodamine 110
or Ala-Ala-Phe-Aminoluciferin)
Dead Cell
Inactive Live-Cell
Protease
100% Live Cells 50% 0% Live / 50 50%
% D De Dead ead ad Cel Cell Cells ls s 100 100%
% Dead DDead
ead Cells
ells
Live Cell
Nucleus
Protease
Active Live-Cell
Protease
Live Cell
Nucleus
Live Cell
Nucleus
Live Cell
Nucleus
Active Dead-Cell
GF+
Protease
Active Dead-Cell
Protease
Live Cell
Nucleus
Active Dead-Cell
Protease
Dead Deaad
d Cell
ell
Dead Cell
Inactive Live-Cell
2
1
0
Live Cell
Active Live-Cell
Protease
Active Dead-Cell
Nucleus
AAF+
Dead Cell
Live
Cells
Protease
Fluorescence at 520nm (rhodamine 110-red) or 505nm (coumarin-blue)
Dead ead Cell ell
Dead
Cells
Overview of the MultiTox-Fluor Multiplex Cytotoxicity Assay protocol.
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Cell Health Assays
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