2013 Promega catalogue

Life
Science
Catalog
2013
Worldwide Contact List
Section
Contents
Cell Signaling
Caspase-Glo ® 3/7 Assay Systems
Product Size Cat.# Price ($)
Caspase-Glo ® 3/7 Assay 2.5 ml G8090 194
For Laboratory Use.
10 ml G8091 661
10 × 10 ml G8093 Pls. Enq.
100 ml G8092 Pls. Enq.
Description: The Caspase-Glo ® 3/7 Assay is a homogeneous luminescent
assay to measure caspase-3/7 activities. The assay provides a proluminescent
caspase-3/7 DEVD-aminoluciferin substrate and a proprietary thermostable
luciferase in a reagent optimized for caspase-3/7 activity, luciferase activity
and cell lysis. Adding the Caspase-Glo ® 3/7 Reagent in an “add-mix-measure”
format results in cell lysis, followed by caspase cleavage of the substrate. This
cleavage liberates free aminoluciferin, which is consumed by the luciferase,
generating a “glow-type” luminescent signal. The signal is proportional to
caspase-3/7 activity. The stabilized luciferase and proprietary buffer system
improve assay performance across a wide range of assay conditions, and the
assay is less likely to be affected by compound interference unlike fluorescentor
colorimetric-based assays. The Caspase-Glo ® 3/7 Assay is designed for use
with multiwell plate formats using either purified enzyme or cells in culture.
Features:
• Simplify Apoptosis or Caspase Detection: The “add-mix-measure”
protocol makes the assay easy to automate; simply add an equal volume of
reagent to sample volume.
• Use Less Enzyme or Fewer Cells: The low background luminescence
results in excellent signal-to-noise ratios and superior sensitivity not
achieved by other caspase formats, allowing assays to be performed in
96- or 384-well formats.
• Decrease Assay Time: No sample preparation or manipulation required,
and no extended incubation times are necessary, as with fluorescencebased
assays. Maximum sensitivity is achieved in as little as 0.25–1 hour.
• Rely on a Performance-Tested Assay: The assay delivers excellent Z´factor
values in cell and purified enzyme models.
• Process Plates in Batch Mode: The extended-glow signal allows the
plates to be read over a 3-hour period of time for batch processing; no
injectors required.
• Get More Information: Multiplex with other cell-based assays from
Promega.
• 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 at –20°C.
Protocol
Caspase-Glo ® 3/7 Assay Technical Bulletin
Part#
TB323
Luminescence (RLU)
Background Subtracted
160,000
12,000
140,000
120,000
100,000
80,000
60,000
40,000
20,000
0
10,000
8,000
6,000
4,000
2,000
0
0 200 400 600 800
0
1,000
2,000
3,000
4,000
5,000
Anti-Fas-Treated Cells
Untreated Cells
Cell #
4098MB05_3A
6,000
7,000
8,000
9,000
10,000
The Caspase-Glo ® 3/7 Assay produces luminescence that is linear
over a broad range of cell numbers. Jurkat cells were treated with
anti-Fas mAb for 4.5 hours to induce apoptosis or were left untreated.
Caspase-Glo ® 3/7 Reagent was added directly to the cells in 96-well
plates and incubated for 1 hour before recording luminescence. Each point
represents the average of 4 wells. The “no cell” blank control value has
been subtracted from each.
Caspase-Glo ® 6 Assay Systems
Product Size Cat.# Price ($)
Caspase-Glo ® 6 Assay 10 ml G0970 655
50 ml G0971 Pls. Enq.
For Laboratory Use.
Description: The Caspase-Glo ® 6 Assay is a homogeneous, luminescent
assay that measures caspase-6 activity. Caspase-6 is a member of the
cysteine aspartic acid-specific protease family and has a key effector role in
the cleavage of specific target proteins during apoptosis. The Caspase-Glo ®
6 Assay provides a luminogenic substrate, Z-VEID-aminoluciferin, in a buffer
optimized for caspase-6 and luciferase activity. The addition of a single
Caspase-Glo ® 6 Reagent in an add-mix-measure format results in cleavage
of the substrate, releasing aminoluciferin, and generation of a glow-type
luminescent signal in the presence of Ultra-Glo Recombinant Luciferase.
The luminescent signal is proportional to the amount of caspase-6 activity
present. The homogeneous Caspase-Glo ® 6 Assay is designed for use with
purified enzyme preparations in multiwell plate formats, making it ideal for
automated high-throughput screening for caspase-6 activity and inhibitors of
caspase-6 activity.
Features:
• Simplified Method: The homogeneous “add-mix-measure” protocol
makes the assay highly amenable to automation.
• Greater Sensitivity: The assay is more sensitive than fluorescence-based
caspase-6 assays. This bioluminescent assay avoids inherent fluorescent
background signals, providing excellent signal-to-noise ratios. The assay is
linear over 3 logs of caspase-6 concentration and can detect 0.002U/ml.
• Increased Accuracy: The superior sensitivity over fluorescence-based
caspase assays allows inhibitor studies at concentrations below the K m .
• Faster Results: The maximum signal (and maximum sensitivity) of the
assay is reached in as little as 30 minutes after reagent addition.
• High-Quality Assay: The assay demonstrates an excellent Z´-factor value
of 0.86 when using 0.1U/ml of caspase-6 for assays in 384-well plates.
• Batch Processing Capability: The coupled-enzyme, homogeneous format
results in a continuous signal, providing excellent stability and allowing
plates to be read over an extended period of time.
Storage Conditions: Store at –20°C.
Protocol
Caspase-Glo ® 6 Assay Technical Bulletin
Part#
TB366
Table of
Contents
38
For complete and up-to-date product information visit: www.promega.com/catalog