2013 Promega catalogue

Cell Signaling
DPPIV-Glo Protease Assay
Product Size Cat.# Price ($)
DPPIV-Glo Protease Assay 10 ml G8350 500
For Laboratory Use.
50 ml G8351 1293
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
DPPIV-Glo Protease Assay Technical Bulletin
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 0.1 1
DPPIV (ng/ml)
Sensitivity of the DPPIV-Glo Protease Assay compared to a
fluorescent assay.
Part#
TB339
4799MA
Proteasome-Glo Assays
Product Size Cat.# Price ($)
Proteasome-Glo Chymotrypsin-Like Assay 10 ml G8621 500
50 ml G8622 1293
Proteasome-Glo Trypsin-Like Assay 10 ml G8631 500
50 ml G8632 1293
Proteasome-Glo Caspase-Like Assay 10 ml G8641 500
50 ml G8642 1293
Proteasome-Glo 3-Substrate System 10 ml G8531 1251
50 ml G8532 Pls. Enq.
For Research Use Only. Not for Use in Diagnostic Procedures.
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
Proteasome-Glo Assay Systems Technical Bulletin
Part#
TB349
7
Drug Discovery
Section
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