2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
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Table of
Contents
Cell Signaling
142
DPPIV-Glo Protease Assay
Product Size Cat.# Price ($)
DPPIV-Glo Protease Assay 10 ml G8350 490.00
For Laboratory Use.
50 ml G8351 1267.00
Description: The DPPIV-Glo Protease Assay is a homogeneous, luminescent
assay that measures dipeptidyl peptidase IV (DPPIV) activity. DPPIV is a serine
protease that cleaves N-terminal dipeptides from polypeptides with L-proline
or L-alanine at the penultimate position. The DPPIV-Glo Assay provides a
proluminescent DPPIV substrate, Gly-Pro-aminoluciferin, in a buffer system
optimized for DPPIV and luciferase activities. The addition of a single
DPPIV-Glo Reagent in an “add-mix-measure” format results in DPPIV
cleavage of the substrate and generation of a “glow-type” luminescent signal
produced by the luciferase reaction. In this homogeneous, coupled-enzyme
format, the signal is proportional to the amount of DPPIV activity present. The
assay is designed for use with purified enzyme preparations.
Features:
• Simplified Method: The homogeneous “add-mix-measure” protocol
makes the assay highly amenable to automation.
• Greater Sensitivity: The assay is more sensitive than fluorescent-based
DPPIV assays. In contrast to fluorescent assays, the luminescent assay
avoids inherent fluorescent background signals and thus provides excellent
signal-to-background readings. The assay is linear over more than 3 logs
of DPPIV concentration and can detect less than 1pg/ml enzyme.
• Faster Results: The maximum signal (and maximum sensitivity) of the
assay is reached in as little as 30 minutes after reagent addition and,
unlike fluorescent assays, is not dependent on accumulation of cleaved
product.
• Amenable to Batch Processing: The stability of the signal allows plates
to be read over an extended period of time.
Storage Conditions: Store components at –20°C protected from light.
Protocol Part#
DPPIV-Glo Protease Assay Technical Bulletin TB339
Signal-to-Noise Ratio
100,000
10,000
1,000
100
10
1
Gly-Pro-aminoluciferin
Gly-Pro-AMC
0.1
0.0001 0.001 0.01
DPPIV (ng/ml)
0.1 1
Sensitivity of the DPPIV-Glo Protease Assay compared to a
fluorescent assay.
4799MA
Proteasome-Glo Assays
Product Size Cat.# Price ($)
Proteasome-Glo Chymotrypsin-Like Assay 10 ml G8621 490.00
50 ml G8622 1267.00
Proteasome-Glo Trypsin-Like Assay 10 ml G8631 490.00
50 ml G8632 1267.00
Proteasome-Glo Caspase-Like Assay 10 ml G8641 490.00
50 ml G8642 1267.00
Proteasome-Glo 3-Substrate System 10 ml G8531 1226.00
For Research Use Only. Not for Use in Diagnostic Procedures.
50 ml G8532 Pls. Enq.
Description: The Proteasome-Glo Assays are homogeneous, luminescent
assays that individually measure the chymotrypsin-like, trypsin-like and
caspase-like protease activities associated with the proteasome in an
enzyme-based format. The 26S proteasome is a 2.5MDa multiprotein complex
found in all eukaryotic cells. Proteasome-Glo Cell-Based Assays provide
luminogenic proteasome substrates in buffers optimized for cell
permeabilization, proteasome activity and luciferase activity. Addition of the
Proteasome-Glo Cell-Based Reagent in an “add-mix-measure” format
results in proteasome cleavage of the substrate and rapid generation of a
luminescent signal produced by the luciferase reaction.
The three luminogenic substrates used to monitor specific protease activities
include: Suc-LLVY-aminoluciferin for chymotrypsin-like, Z-LRR-aminoluciferin
for trypsin-like, and Z-nLPnLD-aminoluciferin for caspase-like activity. Each
luminogenic substrate is added to a buffer system optimized for its specific
proteasome activity and luciferase activity. The reagents are added to test
samples containing proteasome enzyme that cleaves the substrates, releasing
luciferin, which is consumed by luciferase, producing “glow-type” luminescence
correlating to enzyme activity or inhibition.
The Proteasome-Glo 3-Substrate System consists of three
homogeneous bioluminescent assays in an enzyme-based format (each of
these three assays also is available separately).
The Proteasome-Glo Cell-Based 3-Substrate System consists of three
homogeneous bioluminescent assays that measure the three proteolytic
activities associated with the proteasome in a cell-based format (each of these
three assays also is available separately).
Features:
• Simplified Method: The “add-mix-measure” protocol minimizes handling
steps and makes the assays amenable to automation.
• Faster Results: Maximum sensitivity is reached 10–30 minutes after
reagent addition.
• Greater Sensitivity: The luminescent assay format avoids inherent
fluorescent background signals, providing excellent signal-to-background
readings. The assays are miniaturizable to 384-well format.
Storage Conditions: Store the Proteasome-Glo Assay components at
–20°C.
Protocol Part#
Proteasome-Glo Assay Systems Technical Bulletin TB349
For complete and up-to-date product information visit: www.promega.com/catalog