2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
Section
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Table of
Contents
Cell Signaling
Histone Deacetylase Assays
52
HDAC-Glo I/II Assays and Screening
Systems
Product Size Cat.# Price ($)
HDAC-Glo I/II Assay 10 ml G6420 578.00
5 × 10 ml G6421 2137.00
100 ml G6422 2781.00
HDAC-Glo I/II Screening System 10 ml G6430 615.00
5 × 10 ml G6431 2280.00
Product Size Conc. Cat.# Price ($)
HDAC-Glo I/II Control Substrate 10 µl G6550 352.00
HeLa Nuclear Extract 10 µl 5 mg/ml G6570 165.00
Trichostatin A 10 µl 10 mM G6560 142.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description:
The HDAC-Glo I/II Assays and Screening Systems are single-reagentaddition,
homogeneous, luminescent assays that measure the relative activity
of histone deacetylase (HDAC) class I and II enzymes from cells, extracts or
purified enzyme sources. The assays use an acetylated, live-cell-permeant,
luminogenic peptide substrate that can be deacetylated by HDAC activities.
Deacetylation of the peptide aminoluciferin substrate is measured using a
coupled enzymatic system in which a protease in the Developer Reagent
cleaves the deacetylated peptide from the aminoluciferin substrate, releasing
aminoluciferin, which is quantified in a reaction using Ultra-Glo recombinant
firefly luciferase. The assay reaction is typically complete within 15–45 minutes
with no sample manipulation. The HDAC-mediated luminescent signal is
persistent, with a half-life of greater than 3 hours, allowing batch processing of
multiwell plates. The HDAC assay is broadly useful for class I and II enzymes.
The Trichostatin A, included in the HDAC-Glo I/II Screening Systems or available
separately, is a known pan HDAC inhibitor that may be used as a positive
control inhibitor. The Trichostatin A is supplied at a concentration of 10mM in
DMSO.
The HeLa Nuclear Extract, included in the HDAC-Glo I/II Screening Systems
or available separately, may be used as a source of histone deacetylase activity.
The diluted extract also can be used as an HDAC-Glo I/II Assay chemistry
control.
The HDAC-Glo I/II Control Substrate, only available separately, is a nonacetylated
form of the HDAC-Glo I/II Substrate with the same amino acid
sequence and can be used with the HDAC-Glo I/II Assays and Screening
Systems to confirm true HDAC inhibition in secondary screens. The Control
Substrate is supplied at a concentration of 10mM and is sufficient for 480
assays in 96-well plate format when combined with the HDAC-Glo Reagent
prepared with components in the HDAC-Glo I/II Assays or Screening
Systems.
Features:
• Simple Measurement of Deacetylating Activities: Use a singlereagent-addition,
homogeneous, add-mix-measure protocol for easy
implementation from benchtop to screening.
• Highly Sensitive: Obtain 10- to 100-fold higher sensitivity than
comparable fluorescence methods.
• Fast Data Acquisition: Achieve maximum signal in as little as
15 minutes with persistent glow-type steady-state signal, making the
protocol amenable to automation in high-throughput formats and compatible
with luminometers without injectors.
• Flexible to Sample Type: Use with viable cells, extracts or purified
recombinant enzyme sources.
Storage Conditions: Store the HDAC-Glo Assay components and
HDAC-Glo I/II Control Substrate (sold separately) at –20°C. Store HeLa
Nuclear Extract at –70°C.
Protocol Part#
HDAC-Glo I/II Assay and Screening System Technical Manual TM335
Signal-to-Noise Ratio
100,000
10,000
1,000
100
10
1 1
10
100
HDAC Enzyme (ng/ml)
Broad linearity with HDAC Class I and II enzymes.
For complete and up-to-date product information visit: www.promega.com/catalog
1,000
HDAC 1
HDAC 2
HDAC 3
HDAC 8
HDAC 4
HDAC 5
HDAC 7
HDAC 9
HDAC 6
HDAC 10
HDAC 11
8907MA