2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
Section
Contents
Table of
Contents
Cell Signaling
30
Apo-ONE ® Homogeneous Caspase-3/7 Assay
Product Size Cat.# Price ($)
Apo-ONE ® Homogeneous Caspase-3/7 Assay 1 ml G7792 89.00
10 ml G7790 703.00
100 ml G7791 Pls. Enq.
Available Separately
Apo-ONE ® Homogeneous Caspase-3/7 Buffer
For Research Use Only. Not for Use in Diagnostic Procedures.
100 ml G7781 Pls. Enq.
Description: The Apo-ONE ® Homogeneous Caspase-3/7 Assay provides
required reagents for fast and sensitive measurement of active caspase-3 and
-7. The assay includes a profluorescent caspase-3/7 substrate, rhodamine 110
bis-(N-CBZ-l-aspartyl-l-glutamyl-l-valyl-aspartic acid amide; Z-DEVD-R110),
and an optimized cell lysis/activity buffer. The buffer efficiently lyses cultured
mammalian cells and supports optimal caspase-3/7 activity. The substrate and
buffer combine to make the Apo-ONE ® Caspase-3/7 Reagent that is added
directly to samples. Upon cleavage of the DEVD substrate by caspase-3/7, the
rhodamine 110 becomes fluorescent when excited at 498nm (emission maximum
is 521nm). The amount of fluorescent product generated is representative
of the amount of active caspase-3/7 present in the sample.
Features:
• Get Results Faster: The “add-mix-measure” format and high sensitivity
decrease the “time to first result”.
• Use Less Enzyme or Fewer Cells: Optimized caspase-3/7 activity buffer,
in conjunction with the R110-labeled substrate, allow increased sensitivity
over existing fluorescent caspase assay methods.
• Adapt to Your Format and Throughput Needs: Easily onfigure for use
in high-throughput systems and use with purified enzyme preparations, cell
extracts or cultures of adherent, suspension or primary cells.
• Get More Information: Multiplex with other assays: CellTiter-Blue ® Assay
or the Caspase-Glo ® 8 or 9 Assays.
• 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 protected from light and moisture.
Protocol Part#
Apo-ONE ® Homogeneous Caspase-3/7 Assay Technical Bulletin TB295
Fluorescence (RFU)
10,000
8,000
6,000
4,000
2,000
0
0
Z-DEVD-R110 at 1 hour
Ac-DEVD-AMC at 18 hours
10,000
20,000
30,000
Total Jurkat Cells
40,000
Superior sensitivity of the Apo-ONE ® Homogeneous Caspase-3/7
Assay compared to the AMC substrate-based assay.
3396MA05_1A
4028MB
Add an
equal volume
Apo-ONE ®
Reagent
Mix and
incubate
1–18 hours
Record
fluorescence
(498 Ex/521 Em)
Schematic overview of the Apo-ONE ® Homogeneous Caspase-3/7
Assay protocol.
Caspase-3/7 Activity
(RFU x 103 )
26
22
18
14
10
6
TRAIL Caspase-8 Vehicle Caspase-8
TRAIL Caspase-3/7 Vehicle Caspase-3/7
0
1
2
1,200
1,100
1,000
900
800
700
600
500
For complete and up-to-date product information visit: www.promega.com/catalog
3
4
5
Duration of Drug Exposure (Hours)
6
7
8
9
10
Caspase-8 Activity
(RLU)
Multiplexing luminescent Caspase-Glo ® 8 and Apo-ONE ® Caspase-3/7
Assay. The time dependence of caspase-8 and caspase-3/7 activity is
demonstrated.
4754MA