Life Science - Promega

Cell Signaling
CytoTox-Glo Cytotoxicity Assay
Product Size Cat.# Price ($)
CytoTox-Glo Cytotoxicity Assay 10 ml G9290 201.00
For Laboratory Use.
5 × 10 ml G9291 745.00
2 × 50 ml G9292 1115.00
Description: The CytoTox-Glo Assay is a luminescent cytotoxicity assay
that measures the relative number of dead cells in cell populations. The
assay measures the extracellular activity of a distinct intracellular protease
activity (dead-cell protease) when the protease is released from membranecompromised
cells. A luminogenic cell-impermeant peptide substrate (AAFaminoluciferin)
is used to measure dead-cell protease activity. The liberated
aminoluciferin product is measured as “glow type” luminescence generated
by Ultra-Glo Recombinant Luciferase provided in the assay reagent. The
AAF-aminoluciferin substrate cannot cross the intact membrane of viable cells
and does not generate any appreciable signal from the live-cell population.
The amount of luminescence directly correlates with the percentage of cells
undergoing cytotoxic stress. With the addition of a lysis reagent (provided),
the CytoTox-Glo Assay also can deliver the luminescent signal associated
with the total number of cells in each assay well. Viability can be calculated
by subtracting the luminescent dead-cell signal from the total luminescent
value, thus allowing you to normalize assay data to cell number and mitigate
assay interferences that may lead to erroneous conclusions. The cytotoxicity
protease biomarker is constitutive and conserved across cell lines, and the
CytoTox-Glo Assay demonstrates excellent correlation with other methods of
assessing cell viability.
Features:
• Measure the Relative Number of Dead Cells in Culture: Measure
cytotoxicity by adding a single reagent in an “add-mix-measure” protocol.
• Distinguish Between Small Differences in Viability: The assay provides
a linear response and can distinguish between small differences in
viability across the entire spectrum of cytotoxicity, from modest cytotoxicity
(100 to 95% viability) to profound cytotoxicity (5 to 0% viability).
• Normalize Data for Cytotoxicity: Data normalization for dead-cell
number makes results more comparable well-to-well, plate-to-plate and
day-to-day.
• Measure the Relative Number of Remaining Viable Cells Using a
Total Lysis Protocol: Correlate increased cytotoxicity with a reduction in
viable cells.
• Improve your Data: Reduce statistical probability of false-positives (or
false-negatives), and eliminate fluorescence interference issues with a
stable luminescence readout.
H
AAF-N
AAF- +
-
S
N
Dead-cell
protease activity
H 2
N
Luciferase
Mg 2+
S
Light
N
N
S
COOH
N
S
COOH
+ ATP + O 2
Cleavage of the luminogenic AAF-Glo Substrate by dead-cell
protease activity.
Signal to Noise Ratio
2,500
2,000
1,500
1,000
500
0
CytoTox-Glo Assay
Fluorescent LDH Assay
0 2,500 5,000 7,500 10,000
Dead Cells/Well
Superior sensitivity and dynamic range of the CytoTox-Glo Assay
compared to fluorescent LDH Assay.
6802MA
6393MB
2
Cell Health Assays
Storage Conditions: Store at –20°C, protected from light.
Protocol
CytoTox-Glo Cytotoxicity Assay Technical Bulletin
Part#
TB359
Section
Contents
For complete and up-to-date product information visit: www.promega.com/catalog
41
Table of
Contents