EPIGENETICS DRUG DISCOVERY SOLUTIONS

EPIGENETICS DRUG DISCOVERY SOLUTIONS
Optimized for simplicity. Validated for your mark.
Drug Discovery Solutions for Epigenetic and
Other Post-Translational Modifications
• Writers & Erasers of the Histone Code
• p53 Acetylation & Deactylation
Epigenetics, the study of changes in gene expression, is a rapidly growing field which
presents a significant opportunity for drug discovery. As the understanding of the role
of epigenetic changes in various disease states advances, the need for high quality
reagents that can rapidly identify and optimize therapeutic molecules that target these
enzymes is increasing.
To address this need, PerkinElmer has developed first-in-class research tools built upon
our homogeneous assay platforms, LANCE® Ultra and AlphaLISA® technologies. These
platforms offer a combination of specificity, sensitivity, and throughput that other assay
methods lack, all completely validated specifically for drug discovery research applications.
Choose the platform that fits your workflow:
Cellular Detection Kits
In Vitro Tool Box
AlphaLISA & Cell-Histone Buffers rapid and direct detection of
endogenous histone modulation in a
one-well, no-wash assay format
LANCE Ultra – robust, ultra highthroughput TR-FRET technology
AlphaLISA – high sensitivity, high
throughput and ultimate flexibility - assay
short peptides to full-length proteins
PerkinElmer’s suite of epigenetic research reagents offers a faster, more efficient path in the
race to identify promising epigenetic drug candidates:
• Flexibility: Work with peptide or full-length protein substrates, or endogenous histones in
cell-based format
• Sensitivity: Robust and reproducible detection, even at low enzyme concentrations
• Specificity: Reagents highly specific for the mark of interest
• Simplicity: One-well, no-wash assay platforms amenable to automation and HTS
• Validated: Detailed, ready-to-use protocols for over 30 epigenetic enzyme assays
Cellular Detection Kits
The AlphaLISA Epigenetic Cellular Detection Kits enable rapid and
direct detection of endogenous modification of epigenetic marks
on histone H3. These no-wash, all-in-one well assays are
optimized for simplicity and throughput.
• Simple assay protocol amenable to automation, eliminating
tedious ELISAs and Westerns
• Suitable for endogenous and recombinant cell lines, providing
the flexibility to work with relevant cell models
Assay Overview
The AlphaLISA assay is based on a 3-step assay:
cell preparation, extraction and detection*
*Product format, protocol and detection principle for phospho
marks is slightly different – see www.perkinelmer.com/surefire
The detection is based on the immunocapture of the modified
histones on the AlphaLISA beads
ALL-IN-ONE WELL PROTOCOL
PREPARATION
Overnight
EXTRACTION
25 min
DETECTION
90 min
Read Plate
on EnVision®
or EnSpire®
Seed cells in plate
4 hours
Add Cell-Histone
Lysis Buffer
15 min
Add Acceptor Beads and
Biotinylated Anti-H3
60 min
Treat cells –
overnight
Add Cell-Histone
Extraction Buffer
10 min
Add Donor Beads
30 min
Cellular Detection Ordering and Assay Information
Mark Detected
Part Number
Assay Points
Methyl Modifications
Part Number
Assay Points
Normalization Assay
H3K4me2
AL716C500
AL716F5,000
H3K27me3
AL722C500
AL722F5,000
Acetyl Modifications
H3K9ac
AL714C500
AL714F5,000
H3K27ac
AL720C500
AL720F5,000
Phospho Modifications
H3Ser10p*
Mark Detected
TGRH3S500500
TGRH3S10K10,000
TGRH3S50K50,000
AlphaScreen SureFire® GAPDH
Assay Kit*
TGRGD500
500
TGRGD10K
10,000
TGRGDS50K50,000
Additional Items
AlphaScreen® General IgG (Protein A)
Detection Kit
6760617C
500
6760617M
10,000
6760617R50,000
* Requires AlphaScreen General IgG (Protein A) Detection Kit
In Vitro Assay Toolbox
The in vitro toolbox is comprised of validated off-the-shelf reagents for the detection of post-translational modifications of histone H3 and p53.
Validated Enzyme Assays for Histone H3
Full-length
protein assay
available in Alpha
SET7/9
Writers
methylation
Cellular assay
available in Alpha
demethylation
Aurora B
acetylation
P300
HASPIN
PRMT1
PRMT6
deacetylation
P300
G9a
phosporylation
EZH2
NH2 – A R2T3 K4 Q TA R K9 S10 T G G K A P R K Q L AT K A A R K 27 S A PAT G G V K36K P H R . . .
Erasers
JMJD1A
JMJD3
LSD1
JMJD2A
HDAC1
SIRT1
SIRT2
SIRT3
HDAC1
HDAC3/NCOR2
HDAC2
HDAC6
HDAC10
HDAC3/NCOR1
SIRT1
SIRT2
SIRT3
SIRT7
HDAC1
HDAC2
HDAC3/NCOR1
HDAC3/NCOR2
HDAC6
HDAC10
CBP/p300
N – p53 – K382Ac
SIRT1
K
B
+ Enzyme
SAM
AlphaLISA and LANCE Ultra reagents to measure modifications on histone H3 and p53
SAH
Me
All detection reagents have been well characterized for their
specificity at detecting the modification of interest, and validated for use
K
B
with the specific enzymes depicted in the figure above.
+ Eu-Ab
+ ULight-SA
AlphaLISA Beads
LANCE Ultra TR-FRET
FRET
Eu
Em 665 nm
Emission
615 nm
Excitation
320 or 340 nm
Me
ULight
B
Me
K
K
Excitation
680 nm
B
With PerkinElmer, you can count on consistent performance that works every time. Plus we are continuing
to expand our offering of epigenetic tools. If the product you need is not listed please inquire.
In Vitro Ordering and Assay Information
Acetyl Modifications
Validated Assays
Mark Detected Acetyltransferase
Deacetylase
H3K4
SIRT1
unmodified
H3K9/K27
See Note 1
unmodified
H3K9ac
p300 H3K27ac
p300
HDAC1 Alpha Technology
LANCE Ultra (TR-FRET)
AlphaLISA anti-mark Acceptor Beads
LANCE Ultra Europium-labeled anti-mark Antibody
(requires Alpha Streptavidin Donor beads)
(requires LANCE Ultra ULight- Streptavidin)
Part Number
Quantity
Assay Points
Part Number
Quantity
Assay Points
AL119C
250 µg
500
TRF0404-D
10 µg
1,500
AL119M
5 mg
10,000
TRF0404-M
100 µg
15,625
AL119R
25 mg
50,000
AL138C
250 μg 500
TRF0411-D
10 μg 1,500
AL138M
5 mg
10,000
TRF0411-M
100 μg 15,625
AL138R
25 mg
50,000
AL114C
250 µg
500
TRF0400-D
10 µg
1,500
AL114M
5 mg
10,000
TRF0400-M
100 µg
15,625
AL114R
25 mg
50,000
AL120C
250 µg
500
TRF0405-D
10 µg
1,500
AL120M
5 mg
10,000
TRF0405-M
100 µg
15,625
AL120R
25 mg
50,000
Non-Histone Target
p53 K382ac
p300
SIRT1 AL124C
AL124M
AL124R
250 µg
5 mg
25 mg
500
10,000
50,000
TRF0409-D
TRF0409-M
Note 1: For a detailed list of Validated Deacetylase Assays see the “Validated Enzyme Assays for Histone H3” figure above.
10 µg
100 µg
1,500
15,625
Methyl Modifications
Validated Assays
Alpha Technology
LANCE Ultra (TR-FRET)
AlphaLISA anti-mark Acceptor Beads
LANCE Ultra Europium-labeled anti-mark Antibody
Mark Detected Methyltransferase
Demethylase
(requires Alpha Streptavidin Donor beads)
(requires LANCE Ultra ULight™-Streptavidin)
Part Number
Quantity
Assay Points
Part Number
Quantity
Assay Points
H3R2me2
PRMT6
AL139C
250 µg
500
PRMT1
AL139M
5 mg
10,000
AL139R
25 mg
50,000
H3K4
LSD1
AL119C
250 µg
500
TRF0404-D
10 µg
1,500
unmodified
AL119M
5 mg
10,000
TRF0404-M
100 µg
15,625
AL119R
25 mg
50,000
H3K4me1-2
SET7/9
AL116C
250 µg
500
TRF0402-D
10 µg
1,500
AL116M
5 mg
10,000
TRF0402-M
100 µg
15,625
AL116R
25 mg
50,000
H3K9/K27 JMJD1A
AL138C
250 µg
500
TRF0411-D
10 µg
1,500
unmodified
AL138M
5 mg
10,000
TRF0411-M
100 µg
15,625
AL138R
25 mg
50,000
H3K9me2
G9a
JMJD2A
AL117C
250 µg
500
TRF0403-D
10 µg
1,500
JMJD2C
AL117M
5 mg
10,000
TRF0403-M
100 µg
15,625
AL117R
25 mg
50,000
H3K27me2-1
EZH2 JMJD3 AL121C
250 µg
500
TRF0406-D
10 µg
1,500
AL121M
5 mg
10,000
TRF0406-M
100 µg
15,625
AL121R
25 mg
50,000
H3K27me3
G9a AL122C
250 µg
500
TRF0407-D
10 µg
1,500
AL122M
5 mg
10,000
TRF0407-M
100 µg
15,625
AL122R 25 mg
50,000
H3K36me2
JMJD2A AL123C
250 µg
500
TRF0408-D
10 µg
1,500
AL123M
5 mg
10,000
TRF0408-M
100 µg
15,625
AL123R
25 mg
50,000
Phospho Modifications**
Quantity/
Mark Detected
Product Description
Part Number Assay Points
H3Thr3p
LANCE Ultra Europium-labeled anti-phospho-Histone H3 (Thr3) Antibody
TRF0211-D
10 µg / 1,500
LANCE Ultra Europium-labeled anti-phospho-Histone H3 (Ser10) Antibody
H3Ser10p
LANCE Ultra ULight-Histone H3 (Thr3/Ser10) Peptide
TRF0211-M
TRF0210-D
TRF0210-M
TRF0125-D
TRF0125-M
Note
Go to
100 µg / 15,000 www.perkinelmer.
com/kinaseselector
10 µg / 1,500
100 µg / 15,000 to view our kinase
selector tool
0.5 nmole / 1,000
5 nmoles / 15,000
Additional items that need to be ordered with AlphaLISA and LANCE Ultra Methyl / Acetyl assays:
Description
Required for running AlphaScreen Streptavidin Donor beads
AlphaLISA Assays AlphaLISA Biotin-anti-Histone H3 (C-ter) Antibody
(required for full-length histone substrates)
AlphaLISA 5X Epigenetics Buffer 1 Kit
(required for histone H3-derived peptidic substrates)
Required for running LANCE Ultra ULight – Streptavidin
LANCE Ultra Assays
LANCE Detection Buffer, 10 X
Part Number
6760002S
6760002
6760002B
AL118C
AL118M
AL118R
AL008C
AL008F
TRF0102-D
TRF0102-M
TRF0102-R
CR97-100
Quantity
1 mg
5 mg
50 mg
2 µg
40 µg
200 µg
10 mL
100 mL
1 nmole
10 nmoles
100 nmoles
250 mL
*Source of enzyme and substrate provided in detailed protocols available online at: www.perkinelmer.com/epigenetics
**PerkinElmer also offers a wide selection of assays for non-histone phosphorylation events. For more information, visit www.perkinelmer.com/kinase
For more information, please visit www.perkinelmer.com/epigenetics
PerkinElmer, Inc.
940 Winter Street
Waltham, MA 02451 USA
P: (800) 762-4000 or
(+1) 203-925-4602
www.perkinelmer.com
For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs
Copyright ©2011 PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. SureFire is a registered trademark of TGR BioSciences, Pty. Ltd.
AlphaLISA and AlphaScreen reagents are for research use only. All other trademarks are the property of their respective owners.
009997_01
Printed in USA
December 2011
Assay Points
2,000
10,000
100,000
500
10,000
50,000
1,000
10,000
100,000
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