TOOLS FOR EPIGENETICDRUG DISCOVERY| Proteins| Nucleosomes| Antibodies| Assays| Services

DRUG DISCOVERY TOOLBOX
assays, reagents & services to accelerate your research

Epigenetic drug discovery & development programs have attracted much attention and investment in recent years.
This is driven by the potential to develop novel therapeutic strategies that can reverse transcriptional and epigenetic
abnormalities.
Active Motif is the industry leader in providing innovative technologies for epigenetics and gene regulation
research. This brochure features assays, reagents & services that can easily be integrated into your researchstrategy to advance and streamline the drug discovery workflow.

Assay-ready Enzymes

TARGET
IDENTIFICATION

Histone Substrates
Recombinant Nucleosomes
Small Molecules

SCREENING

Target-relevant Antibodies
Histone Peptide Array
Multiplex Epigenetic Assays

LEAD
OPTIMIZATION

Global Response Assays
Genome-wide Services
Pathway Analysis

DRUG
DISCOVERY

miRNA Functional Assays
Screening Tools
PRECLINICAL/
CLINICAL TRIALSFor a complete list of available products, please visit us at www.activemotif.com.
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Tools for Drug Discovery
proteins, substrates, antibodies & assays for epigenetics drug discovery
Epigenetic targets are the most promising class of druggable targets to emerge in a decade. These targets are not
only relevant to oncology research, but they also have potential impact on metabolic, neurological, inflammatory
and cardiovascular disorders. Active Motif offers everything from highly pure, validated antibodies and proteins to fully
developed assays and services to help advance research in all phases of drug discovery.
> 700 VALIDATED ANTIBODIES

Small
molecule
Epigenetic
Modulators

Drug
Screening

RecombinantProteins

TargetVerification> 300 PURIFIED ASSAY-READY PROTEINS• Bromodomains• Chromatin modifiers• Methyltransferases• Demethylases• Acetyltransferases• Deacetylases• Transcription factors• Nuclear receptors

Pre-clinical
Tests

Histones &Nucleosomes

HATs and HDACs

Histone Modifiers

HMTs & HDMs

Bromo Domains

Ubiquitination

DNA Methylation
Transcription
Factors
Kinases

DNA Damage

Active motif offers a large collection of high quality, robust epigenetic
modifying enzymes and substrates that are ideally suited for use
in drug discovery for the development of activity assays for drug
screening applications.

IDENTIFY MECHANISM OF ACTION AT THE EPIGENETIC LEVELHistone Substrates

PHYSIOLOGICALLY-RELEVANT ASSAY SUBSTRATES

istone Substrates

ctamerTargetDiscovery• Histone modifications• Chromatin modifiers• DNA methylation• DNA damage• Transcription regulation• Stem cell
• Nuclear receptors• Signal transduction

PeptidesHistones

Mono
nucleosome

Octamer(Di, Tri …) Oligo
Nucleosomes

Mononucleosome(Di, Tri …) Oligo
Nucleosomes

Active Motif offers a suite of Services specifically designed toenhance drug development programs, especially those focusedon small molecule epigenetic inhibitors. In the early phases ofdrug discovery, our recombinant proteins and cell-based screening
services will accelerate your program, while in the late phaseour genomic assays, such as ChIP-Seq, RNA-Seq, MeDIP-Seq andOpen Chromatin-Seq will lead to a greater understanding ofyour compound’s mechanism of action.

For a complete list of available purified recombinant proteins, please visit us at www.activemotif.com/proteins.
sales@activemotif.com

1

Epigenetic Proteinshighly purified recombinant proteins for epigenetics research
One of the current challenges in the field of epigenetics research and drug discovery is developing robust enzymatic
assays. The key to achieving a robust enzymatic assay is the ability to obtain high quality proteins.
Active Motif has developed a large selection of full-length high quality, robust epigenetic modifying enzymes for use in thedevelopment of activity assays for drug discovery. Our offerings include N-terminal tagged versions as well as several
protein complexes, including PRC2 complex and MLL complex. Our proteins have been tested in a variety of highly
sensitive enzyme activity assay technologies, including AlphaLISA, HTRF, fluorography and mass spectrometry.
> 360 PROTEINS AVAILABLE

Purification of Full-length Recombinant Proteins• Full length proteins and catalytic domains• Tagged proteins• Protein complexes• Custom and bulk production• Activity by HTRF and AlphaLISA• Purity by SDS-PAGE

KAT5

HDAC6

kDa
170
130
100

READERSWRITERSERASERS• BRD1• BRD4• BRD9• BAZ1B• SMARCA4• TRIM24• PRMT• p300• EZH2• DOT• SET• SMYD• HDAC• JMJ• LSD1• KDM5A/B
• UTX
• TET

kDa

170
130

130

100

100

170

70

PRC2 Complex
130
SUZ12

55

EZH2

170
130

EED

100

RbAp48
RbAp46

40

40

LSD1
kDa

kDa

70

55

55

40

100

70

70

55

TET1-CDkDa

BRD4 (44-168)
kDa
55
40
35
25
15

70
55

Figure 2: Full-length recombinant proteins and protein complexes
expressed and purified. The Coomassie stains reveal that recovered
proteins are full length with little or no detectable degradation anddetermined to have ≥ 98% purity.

Figure 1: Recombinant BAZ1B (1340-1457) HTRF activity assay.
3.3 µM histone peptide H4K5/8/12/16(4Ac) was incubated with BAZ1B(1340-1457) protein in reaction buffer including 50 mM HEPES-NaOH
pH 7.0, 0.1% BSA overnight at room temperature. Anti-FLAG antibody
was used to detect reaction products.

IN-HOUSE MANUFACTURED AND QC’D
• Custom & bulk orders available• Please contact us atwww.activemotif.com/epiproteins

For a complete list of available purified recombinant proteins, please visit us at www.activemotif.com/proteins.
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Bromodomain Proteins
the most comprehensive reader domain collection available for drug discovery
Bromodomain proteins play an integral role in the regulation of transcription and chromatin remodeling. Because
bromodomain proteins have been shown to also regulate transcription of certain oncogenes, they are promisingtherapeutic targets for cancer treatment. To support research efforts, Active Motif offers a comprehensive collectionof recombinant bromodomains for drug discovery research. Our recombinant proteins are manufactured to meetthe highest standards of purity and activity required for conventional drug discovery assays, such as HTRF (Figure 1),
mass spectrometry, AlphaLISA and Alphascreen. These assays are routinely incorporated as part of our quality control
process to ensure that our recombinant proteins and enzymes demonstrate the nM range activities needed for use in
identification of potent inhibitors.

DESIGNED SPECIFICALLY FOR USE IN ASSAYS AND SCREENS

BRD4 (44-168) Titration

15K

BRD4
I-BET
Ac

Transcription

Ac

I-BET

Ac

10K
HTRF Signal

BRD4

Ac

5K

Inflammatory genes

Reduced inflammatory response

0

ACTIVE MOTIF BROMODOMAIN PROTEINS

0

BRD1 (556-688)

31438

BRD3 (24-144)

31379

BRD4 (44-168)

31380

BRD7 (129-236)

31381

150

200

250

300

250

300

BRD4 (333-460) Titration

15K

10K
HTRF Signal

Catalog No.

100

Bromodomain conc. (nM)

• Over 40 reader domains available• Designed for epigenetic drug discovery• Tagged versions available• Lot specific data, including:- activity by HTRF and AlphaLISA- purity by SDS-PAGE

Product

50

5K

0
0

50

100

150

200

Bromodomain conc. (nM)

BRD9 (130-259)

31382

BRDT (21-137)

31450

BRPF1 (627-746)

31375

CREBBP (1081-1187)

31373

SMARCA2 / BRM (1367-1511)

31449

activity were as follows: 3.3 µM histone peptide H4K5/8/12/16(tetra-acetyl)

TAF1 (1398-1524)

31403

50 mM HEPES-NaOH pH 7.0, 0.1% BSA for 1 hour at room temperature.

TRIM24 (862-980)

31368

Anti-FLAG antibody was used to detect the reaction products.

Figure 1: BRD4 (44-168) (top panel), and BRD4 (333-460) (bottom panel)bromodomains tested by HTRF. Assay conditions for bromodomain (BRD)
was incubated with the protein indicated in reaction buffer containing

For a complete list of available purified recombinant proteins, please visit us at www.activemotif.com/proteins.
sales@activemotif.com

3

Methyltransferases and Demethylases
active, recombinant HMT and HDM proteins

H3K4me3

4700 Reflector Spec #1 [BP = 713.4, 59817]

1007.5100

No Enzyme[H3K4me3] = 3.3 µm% Intensity

Active Motif has produced over 60 methyltransferases
and demethylases as well as several enzymatic complexes
for many relevant drug targets, including NSD2, DOT1L, LSD1,KDM5A, KDM5B, KDM6A, and PRC2 and MLL complexes.
The collection also includes N-terminal FLAG-tagged
versions of HMTs and HDMs as well as a number of
DNA methyltransferases (DNMTs).

MALDI-TOF Analysis Reveals High Level of Demethylase
Activity of KDM5A / JARID1A Recombinant Protein

80
60
40
20
0
2690

2706

2698

2714

2722

2730

Mass (m/z)

4700 Reflector Spec #1 [BP = 713.4, 51764]
100[KDM5A] = 100 nm[H3K4me3] = 3.3 µm% Intensity

One of the current challenges in the field of epigenetics
research and drug discovery is developing robust enzymatic
assays. The key to achieving a robust enzymatic assay is theability to obtain high quality proteins.

633.3

H3K4me280
H3K4me160

H3K4me3

40
20
0
2690

Our HMTs and HDMs are manufactured to meet the high­est
standards of purity and activity required for drug dis­covery
assays including fluorography, mass spectrometry, AlphaLISA
and TR-FRET. In addition, dose response assess­ments
performed with various reference compounds ensure
utility of the enzymes in inhibitor studies.

2706

2698

2714

2722

2730

Mass (m/z)

Figure 1: Mass spectrometry data demonstrate that our recombinant KDM5A enzyme is highlyrobust, requiring extremely low nM range amounts of protein to achieve high catalytic rates.
The data show that only 100 nM KDM5A is sufficient to convert 65% 3.3 µM H3K4me3 peptides
to H3K4me2 and 25% 3.3 µM H3K4me2 peptides to H3K4me1 in 1 hour. The catalytic rate of
KDM5A: 40 turnovers/enzyme molecule/hour.

Activity Test for KDM5A using TR-FRET
SPECIFICALLY DESIGNED FOR EPIGENETICDRUG DISCOVERY

20K
HTRF Signal• Custom & bulk orders available• Tagged versions available• Lot specific data, including:- activity by HTRF and AlphaLISA- purity by SDS-PAGE and mass spec

KDM5A Titration

25K

15K
10K
5K
0

0

20

40

60

80

100

120

Enzyme Conc. (nM)

Z Factor of KDM5A Assay
Product

20K

Catalog No.

S/N=14, Z´=0.87

15K

31474

EZH2 Complex

31337

JARID1A / KDM5A

31431

JARID1B / KDM5B

31432

LSD1 / KDM1A

31426

MLL1 Complex

31423

Wells

NSD1-SET

31475

Figure 2: Date generated for the development of an HTRF (Homogeneous Time Resolved Flu-

PRC2 EZH2 (Y641C) Complex

31389

PRMT5

31393

SET2

31358

SMYD2

31497

HTRF Signal

DOT1L (1-416)

10K

0% Inhibition
100% Inhibition (no enzyme)

5K
0

0

4

8

12

16

20

24

28

32

orescence) assay in cooperation with Cisbio. HTRF combines fluorescence resonance energy
transfer technology (FRET) with time-resolved measurement (TR) and is the most common assay used in drug discovery for high-throughput screening (HTS) of drug targets. Titration curve(left) was generated to show signal response in the presence of modified peptide substrate atincreasing protein concentrations. The Z-factor assessment (right) was performed to quantify.
the suitability of assay for use in a full-scale, high-throughput screen.

For a complete list of available purified recombinant proteins, please visit us at www.activemotif.com/proteins.
4

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HATs and HDACS
acetyltransferases and deacetylases
HATs and HDAcs play an important role in gene regulation. Changes in acetylation patterns are observed between
normal cells and cancer cells. In recent years, histone acetylases and deacetylases (HATS & HDACs) have emerged asimportant drug target candidates.
Active motif offers a large selection of HAT and HDAC proteins (full length and catalytic domains) as well as antibodies,
inhibitors and fully validated kits for use in drug screening assays. Our recombinant proteins are manufactured to meetthe highest standards of activity (Figure 1) and purity (Figure 2) required for conventional drug discovery assays.
14 HDACs AND HATS

Purity by SDS-PAGE• Full-length proteins and protein complexes• Highly active• Purity by SDS-PAGE• Custom and bulk orders available

HATs & HDACs
KAT5

KAT7

kDa

kDa

170

170

130

130

100

100

70

70

55

55

40

40

HDAC1
kDa
250
150
100
Figure 1: Recombinant p300 (1041-1161) activity assay. 3.3 µM histone peptide

75

H4K5/8/12/16(4Ac) was incubated with p300 (1041-1161) protein in reaction
buffer including 50 mM HEPES-NaOH pH 7.0, 0.1% BSA for 1 hour at room tem-

50

perature. Anti-FLAG antibody was used to detect reaction products.

37

Product

Catalog No.

HDAC1

31342

HDAC3 Complex

31349

HDAC4

31350

HDAC5-catalytic domain

31351

HDAC6

31346

HDAC7

31352

HDAC8

31353

HDAC9

31354

p300

31124

SIRT1

31340

SIRT6

31336

HDAC6
kDa
170
130
100
70
55

Figure 2: Full-length recombinant proteins and protein complexes
were expressed and purified. Purified recombinant proteins wererun on an SDS-PAGE gel and stained with Coomassie blue. The
Coomassie stains reveal that recovered proteins are full length withlittle or no detectable degradation and were determined to have≥ than 98% purity.

IN-HOUSE MANUFACTURED AND QC’D
• Custom & bulk orders available• Please contact us atwww.activemotif.com/epiproteins

For a complete list of available purified recombinant proteins, please visit us at www.activemotif.com/proteins.
sales@activemotif.com

5

DNA Methylation Proteins
DNA methyltransferases and TET family
Changes in the methylation state have been implicated in many malignancies. As a result, DNA methyltransferase enzymes(DNMTs) have emerged as attractive therapeutic targets. Several therapeutic agents that are DNMT inhibitors have alreadybeen identified and there are many active programs searching for new inhibitory molecules. Other proteins involved in theregulation of DNA methylation include the TET family of proteins which have emerged as potential druggable targets.
Active Motif offers a range of DNA methylation-related proteins for use in drug screening including DNMTs and TET proteins.
Our recombinant proteins are manufactured to meet the highest standards of purity and activity required for conventional
drug discovery assays. In addition we offer a range of validated antibodies to proteins involved in the methylation process aswell as against methylcytosine variants such as 5-mC, 5-hmC, 5-fC and 5-caC.

Figure 1: Recombinant DNMT3A protein activity assay. Recombinant DNMT3A protein

Figure 2: SF9 expressed DNMT proteins. Coomassie staining

activity was measured using the Active Motif DNMT Activity / Inhibition Assay (Catalog

reveals that recovered proteins are full-length with little or no

No. 55006), for 1 hour at 37°C.

detectable degradation and determined to have greaterthan or equal to 98% purity.

Product

Catalog No.

DNMT1

31404

DNMT3A

31406

DNMT3A / DNMT3L Complex

31415

DNMT3B

31413

ACTIVE MOTIF DNA METHYLATION PROTEINSDNMT3B / DNMT3L Complex

31416

DNMT3L

31414

TET1 (1418-2136)

31417

TET2 (1129-2002)

31418

TET3 (824-1795)

31421• Specifically designed for epigenetic drug
discovery• Lot specific data, including:- activity data
- purity by SDS-PAGE

For a complete list of available purified recombinant proteins, please visit us at www.activemotif.com/proteins.
6

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Substrates For Epigenetic Enzymes
physiologically relevant substrates for biochemical assays
A challenge associated with epigenetic screening assays is the choice of substrate. The biological targets of the epigenetic
enzymes within the cell are not free histones. Rather, histone targets are organized into nucleosomes that are further packaged into high order structures within chromatin. The complexity and structural variability of chromatin represents a uniquedrug discovery challenge for these classes of enzymes because it is difficult to reproduce the chromatin structure in vitro.
Therefore, alternative substrates, such as short modified peptides that can effectively mimic the histone tail, are often used inhigh-throughput screens.
However, peptides do not reproduce the complexity of the native biological substrate and do not always interact with
epigenetic enzymes in a way that yields a productive reaction. Other types of substrates have been shown to perform betterwith specific enzymes and include modified or unmodified full-length histones, histone octamers, mononucleosomes andoligonucleosomes (Figure 1). Therefore, critical decisions need to be made during assay development with regards to whatsubstrate choice will perform best with its associated enzyme in the assay.
Mononucleosome H3.1

LARGEST COLLECTION OF SUBSTRATESHistone H3.1
H3.1 Peptide• Nucleosomes
SAH (nM)• Histone octamers• Biotinylated substrates

SAH (nM)

1500• Unmodified and modified histones

1000

Histone Substrates
500

0

0

50

100

150

200

PRC2 (nM)tone Substrates
A

amer

Octamer H3.1

A

2000

2000

PeptidesOligonucleosome H3.1

Mononucleosome H3.1
Octamer H3.1

Histones

Histone H3.1
H3.1 Peptide

Mononucleosome

Octamer

1000

Mononucleosome(Di, Tri …) Oligo
Nucleosomes

500

0

50

100

150

IN-HOUSE MANUFACTURED
PRC2 (nM)AND QC’D
• Custom & bulk orders available• Please contact us atwww.activemotif.com/epiproteins

SAH (nM)

SAH (nM)

SAH (nM)

Histone H3.1
B(Di, Tri …) Oligo
H3.1 Peptide
Nucleosomes
Histone H4

600

1500

0

Octamer H3.1

800

400

200

200

0
200

400

600

800

1000

SETD2 (nM)

Figure A: PCR2 favors mononucleosomes over other substrate alternatives. Activity of
PRC2, a complex of EZH2, EED, SUZ12 and RbAp46/48, towards a variety of substrates
was measured using an HTRF assay detecting conversion of SAM to SAH.
Figure B: SETD2 has an absolute requirement for nucleosome substrates. Comparisonof SETD2 activity towards a variety of substrates measured using an HTRF assay.

For a complete list of available substrates, please visit us at www.activemotif.com/proteins.
sales@activemotif.com

7Full-Length Histones
unmodified & modified histone substrates
Active Motif offers an exclusive collection of over 60 full-length
unmodified and modified recombinant histones, includingsite- and degree-specific modifications such as methylation,acetylation and phosphorylation. Because of our patented
Expressed Protein Ligation (EPL) and Methylated Lysine Analog(MLA) synthesis technologies, Active Motif is the exclusive
source of most of our full-length modified histones.
The outcome of your in vitro biochemical assay is moreaccurate when the system mimics actual cellular biology. Use
our histone proteins as stand-alone substrates or assemble togenerate nucleosomes and oligonucleosomes. Alternatively,
save time and effort by choosing from our selection of
pre-assembled human histone octamers and nucleosomes(see opposite page).

THE LARGEST COLLECTION OF
RECOMBINANT HISTONES AVAILABLE• Over 60 recombinant histones• Site- and degree-specific modifications• Patented synthesis technologies• Biotinylated histones available for FRET

Product

WHICH HISTONE SUBSTRATE IS RIGHT FOR YOUR ASSAY?

Figure: Choosing the correct histone substrate for assay development is key to achieving the best results. Opinion leaders in epigenetics recognize the power of reconstituting recombinant chromatin for creating biologically relevant substrates.

Catalog No.

Histone H3.1 biotinylated

31296

Histone H2A

31490

Histone H2B

31492

Histone H431223

Histone H3K4me1

31208

Histone H3K4me2

31277

Histone H3K4me3

31278

Histone H3K9me1

31281

Histone H3K9me2

31280

Histone H3K9me3

31279

Histone H3K9ac

31253

Histone H3K14ac

31254

Histone H3K18ac

31273

Histone H3K27ac

31290

Histone H3K27me3

31216

Histone H3K36me2

31218

Histone H3K79me2

31221

Histone H4K20me1

31224

For a complete list of available recombinant histones, please visit us at www.activemotif.com/recombhis.
8

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s

Nucleosomes and Histone Octamers
physiologically relevant assay substrates
ACHIEVE BETTER RESULTS WITH
THE RIGHT CHOICE OF SUBSTRATE

Whether performing drug screens, analyzing enzyme
kinetics or monitoring changes in histone modifications,substrate selection is critical.
Pre-assembled histone octamers and nucleosomes
better mimic cellular biology when compared to histone
proteins alone or synthetic peptides.

1

3

4 5

NSD1-SET1

ADVANTAGES

2

2

3

4 5

NSD2-SET• More physiologically relevant• Improves enzyme activity• Better assay specificity• Biotinylated versions available

1. No enzyme
2. No substrate
3. Histone Octamers
4. Nucleosomes A
5. Nucleosomes B

Figure 1: To illustrate the importance of choosing the correct substrate toachieve the best results, recombinant nucleosomes were compared with histone
octamers in a histone methyltransferase assay using SET domain-containing HMT
enzymes MMSET/NSD1-SET and NSD2-SET. The results show there is greater methyltransferase activity when using recombinant nucleosomes as substrates when

In vivo, histones are assembled into octamers andare wound by DNA in chromatin, making octamersand nucleosomes more physiologically relevant
substrates than histones and histone-derived peptides
for in vitro studies. More importantly, some histone
methyltransferases (HMT), such as DOT1L and NSD
enzymes, are significantly more active and specific whenusing nucleosome substrates in HMT assays. Nucleosomes
are also widely used in HMT screening assays to identify
small molecule inhibitors for drug discovery.
No assembly required
Active Motif’s recombinant histone octamers andnucleosomes come pre-assembled to save you timeand money. Histone octamers are comprised ofunmodified core histone proteins (H2A, H2B, H3 and H4).
For nucleosomes, the histone octamer is bound by DNA.
Both unlabeled and biotin-labeled versions are available
to enable flexibility in your experimental design.

Histone Substrates

OctamerMononucleosome(Di, Tri …) Oligo
Nucleosomescompared to octamers alone.

Figure 2: Recombinant nucleosomes(H3.1) run on an SDS-PAGE gel andstained with Coomassie blue.

Product

Catalog No.

Histone Octamer (H3.1)

31470

Histone Octamer (H3.1) - biotinylated

31471

Histone Octamer (H3.3)

31472

Histone Octamer (H3.3) - biotinylated

31473

Nucleosomes (H3.1)

31466

Nucleosomes (H3.1) - biotinylated

31467

Nucleosomes (H3.3)

31468

Mononucleosomes (H3.3) - biotinylated

31469

Nucleosome Preparation Kit

53504

Chromatin Assembly Kit

53500

Nucleosome Assembly Control DNA

53502

For a complete list of available substrates, please visit us at www.activemotif.com/proteins.
sales@activemotif.com

9Small Molecules
to modulate activity of epigenetic enzymes
Active Motif offers an expanding collection of small molecule compounds (activators and inhibitors) to modulate theactivity of proteins that regulate DNA methylation, chromatin and histones.
These compounds are ready for use as epigenetic drug discovery tools for lead generation and assay development.
Our comprehensive selection includes compounds that target DNA methyltransferases (DNMTs), histone acetyltransferases(HATs), histone deacetylases (HDACs), histone methyltransferases (HMTs), histone demethy­lases (HDMs), and the BET
family of bromodomains. These products can be readily scaled to adapt to the requirements of your research.
Description

TargetCat. No.

LYSINE DEMETHYLASE AND METHYLTRANSFERASE

Description

TargetCat. No.

DNA METHYLATION

AS-8351

KDM5B inhibitor

14112

2´-Deoxy-5-fluorocytidine DNMT inhibitor

14108

BIX-01294

G9a inhibitor

14072

14100

Daminozide

KDM2/KDM7 inhibitor

14058

Decitabine
DNMT inhibitor(5-Aza-2´-deoxycytidine)DMOG

PHD/JMJD2A inhibitor

14062

5-Azacytidine

DNMT inhibitor

14102

6-Thioguanine

Degradation of DNMT

14126

Mithramycin A

DNMT1 inhibitor

14128

RG108

Non-nucleoside DNMT
inhibitor

14104

Zebularine

DNMT inhibitor

14106

GSK-J1 (cell impermeable) JMJD3/UTX inhibitor

14068

GSK-J4 (cell permeable) JMJD3/UTX inhibitor

14070

IOX1

JMJD family inhibitor

14056

ML-324

JMJ2 inhibitor

14078

PBIT

JARID1 inhibitor

14110

Tranylcypromine
hemisulfate

LSD1/BHC110 inhibitor

14046

SIRTUIN
AK-7

SIRT2 inhibitor

14054

Chaetocin

Su(VAR)3-9 inhibitor

14051

BML-278

SIRT1 activator

14024

NSC-663284

SETD8 inhibitor

14122

EX-527

SIRT1 inhibitor

14028

UNC-1999

EZH2 inhibitor

14114

Piceatannol

SIRT1 activator

14036

Resveratrol

SIRT1 activator

14022

Salermide

SIRT1/SIRT2 inhibitor

14124

SirtAct

SIRT1 activator

14080

SirtinolSirtuin inhibitor

14074

Splitomicin

Sir2p (yeast Sirt1
homolog) inhibitor

14086

Triacetyl-resveratrol

Sirtuin activator

14116

JQ1 (racemic)

BET bromodomain
inhibitor

14066

RVX-208

BET bromodomain
antagonist

14090

HISTONE ACETYLTRANSFERASE
C646

p300/CBP inhibitor

14052

CPTH2

GCN5P inhibitor

14094

CTPB

p300 activator

14064

Embelin

PCAF inhibitor

14118

Garcinolp300/PCAF inhibitor

14076

Apicidin

HDAC inhibitor

14040

BML-210HDAC inhibitor

14048

CI-994HDAC inhibitor

14092

CUDC-101

HDAC & receptor tyrosine kinase inhibitor

14060

HISTONE DEACETYLASEBROMODOMAIN

ARGININE METHYLTRANSFERASEHPA (Hexyl-4-pentynoic acid) HDAC inhibitor

14034

MS-275HDAC inhibitor

14042

Panobinostat

Class I & II HDAC inhib.

14044

OTHER

Phenylbutyrate Na

HDAC inhibitor

14033

Romidepsin

HDAC inhibitor

14083

TM-2-51

HDAC8 activator

14096

Trichostatin A

HDAC inhibitor

14038

Tubastatin A hydrochloride HDAC6 inhibitor

14084

Valproic acid, sodium salt HDAC inhibitor

14021

Vorinostat (SAHA)

14026

HDAC inhibitor

TC-E-5003

PRMT1 inhibitor

14099

Sinefungin

Methyltransferase (DNA,RNA, protein) inhibitor

14088

UNC-2170

Methyl-lysine binding
protein inhibitor

14120

For a complete list of available activators and inhibitors, please visit us at www.activemotif.com/smallmol.
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Antibodies
for discovery & analysis of druggable targets
Active Motif is more than just an antibody supplier. We
identify, manufacture and rigorously test each antibody in
our vast collection to ensure that we offer only the highest
quality product.
We monitor every step from immunogen design andspecificity screening to application validation. This thoroughprocess ensures our antibodies meet the high standards forconsistency and quality you require to achieve the goals ofyour drug discovery and development efforts.
ANTIBODIES FOR RELEVANT DISEASE AREAS• Oncology• Immunology• Inflammation• Neurobiology• Metabolic disorders• Cardiovascular disease
OVER 700 KEY DNA METHYLATION, HISTONE AND
TRANSCRIPTION FACTOR TARGETS

EPIGENETICS

TRANSCRIPTION FACTORS• H2A, H2B, H3 & H4 PTMs
• DNA methylation• Bromodomain proteins• Methyltransferases• Demethylases• Acetyltransferases• Deacetylases• Hormone receptors• NFkB family• STAT family• IRF family• AP-1 family• p53• Oct4

NEW• Recombinant antibodies to epigenetic marks• Custom conversion of monoclonals• Proprietary tag for enzymatic labeling andsolid support attachment• Please contact us for more information

ADVANTAGES• Rigorous application testing• Assay-compatible formulations• Large lot sizes• Monoclonals to histone PTMs

Product

Catalog No.

BRD4 antibody (pAb)

39909

DOT1L antibody (pAb)39953

EED antibody (mAb)

61203

EZH2 antibody (mAb)

39875

HDAC1 antibody (mAb)39531

HDAC2 antibody (mAb)39533

Histone H3K4me3 antibody (mAb)

61379

Histone H3K27me3 antibody (mAb)

61017

Histone H3K36me2 antibody (mAb)

61019

Histone H3K79me2 antibody (pAb)39143

LSD1 / KDM1A antibody (pAb)

61607

MMSET / WHSC1 antibody (mAb)39879

Smyd3 antibody (pAb)

61407

For more information and a complete list of available antibodies, please visit us at www.activemotif.com/abs.
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11

Screen Binding Specificity
of antibodies, proteins & enzymes
The MODified™ Histone Peptide Array enables rapid,
high-throughput screening of antibodies, enzymes andproteins for cross-reactivity or binding interactions withhistones and their modifications. Each peptide array
contains 384 unique histone modification combinations
in duplicate, with up to four separate modifications on
the same 19mer peptide.

Anti-H3K9me3; anti-Rabbit IRDye 800• Most extensive coverage of modifications on the market
384 unique combinations enable the study of not
only individual sites, but also neighboring effects ofmodifications on recognition and binding.

Anti-H3K27me2me3; anti-Mouse IRDye 680• Unique synthesis & conjugation methods
ensure >95% purity of bound peptides and high peptide
density at each spot, which is advantageous for analysis
of interaction sites with low binding constants.
• Simple Western blot-like array protocol
no special equipment needed.

H3K9me3; H3K27me2me3

Two Array Labeling Kit versions are available:• MODified Array Labeling Kit
to study antibody and enzyme interactions.
• MODified Protein Domain Binding Kit
to analyze the binding specificity of His-tagged
protein reader domains.

Figure: Analysis of the MODified Histone Peptide Array using the LI-COR Odyssey Infrared Imaging System. The top image shows the binding pattern of theH3K9me3 antibody alone, the middle image shows the binding pattern of theH3K27me2me3 antibody in combination with the c-Myc control antibody from

Product

the Array Labeling Kit, and the bottom image is the composite from the Odyssey

Catalog No.

MODified™ Histone Peptide Array, 1 array

13001

MODified™ Histone Peptide Array, 5 arrays

13005

MODified™ Array Labeling Kit

13006

MODified™ Protein Domain Binding Kit

13007

system. The results of the LI-COR Odyssey Infrared Imaging system show that dual
detection is possible with the MODified Histone Peptide Arrays.

Application

Array

Labeling Kit

Antibody Specificity

MODified™ Histone Peptide Array

Array Labeling Kit

Enzymatic Effects

MODified™ Histone Peptide Array

Array Labeling Kit

Protein Domain Specificity

MODified™ Histone Peptide Array

Protein Domain Binding Kit

Table of MODified array reagents recommended for specific applications. The MODified Array Labeling Kit and the MODified Protein Domain Binding Kit are complementary products
containing the buffers and reagents required to process the array for detection. The above table shows the combination of reagents recommended for each application.

For more information on the MODified Histone Peptide Array and Labeling Kits,
please visit us at www.activemotif.com/modified.
12

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Measure Global Changes in the Epigenome
generate histone profiles of your cell models
Active Motif offers a variety of high-throughput, quantita­
tive, ELISA-based assays to measure global changes inhistone modifications in response to drug treatment.

MEASURE CHANGES IN HISTONE MODIFICATIONSIN RESPONSE TO DRUGS

Screen the effects of compounds and other variables
on histone modification levels
The addition or removal of modifications such as phospho,
methyl and acetyl functional groups to histones have a
profound effect on the regulation of transcription, chromosome packaging and DNA damage repair. Screening
extracts for specific histone modifications is a simple way
to assess cell health and compound effects.
Active Motif’s comprehensive selection of Histone Modification ELISAs provides a simple solution for screeningchanges in histone H3 modification levels from purified
core histones (see Product Ordering Information for a listof available Histone Purification Kits) or histones isolatedby acid extraction.

Figure: Histone H3 acetyl Lys27 (H3K27ac) ELISA. The Histone H3 acetyl Lys27 ELISA was used
to assay HeLa acid extracts (5 μg) that were either untreated (–) or treated with sodium
butyrate (+), a known HDAC inhibitor. The provided Recombinant Histone H3 acetyl Lys27

HISTONE MODIFICATION ELISAS FOR:
• Total Histone• H3K4me1• H3K4me2• H3K4me3• H3K9me2• H3K9me3• H3K27me1• H3K27me3• H3K9ac• H3K14ac• H3S10ph• H3S28ph

ASSAY FEATURES

protein was assayed from 0.67 - 3.75 ng/well as a reference standard curve.

Product

Catalog No.

Total Histone H3 ELISA

53110

Histone H3 monomethyl Lys4 ELISA

53101

Histone H3 dimethyl Lys4 ELISA

53112

Histone H3 trimethyl Lys4 ELISA

53113

Histone H3 acetyl Lys9 ELISA

53114

Histone H3 dimethyl Lys9 ELISA

53108

Histone H3 trimethyl Lys9 ELISA

53109

Histone H3 phospho Ser10 ELISA

53111• QUANTITATIVE – Includes control standards for easy
and accurate quantitation

Histone H3 acetyl Lys14 ELISA

53115• THROUGHPUT – Stripwell microplate format for
manual or high throughput analysis

Histone H3 monomethyl Lys27 ELISA

53104

Histone H3 trimethyl Lys27 ELISA

53106

Histone H3 phospho Ser28 ELISA

53100

Histone Purification Kit

40025

Histone Purification Mini Kit

40026

Histone Purification Microplate Kit

40027• SIMPLE – ELISA based colorimetric assay• SPECIFIC – tested for cross-reactivity with other
degree-specific histone modifications

To learn more about our histone analysis products, visit www.activemotif.com/hismodinfo.
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13

Measure Global Changes in the Epigenome
assay specific & off-target effects in a single well

H3

Histonecontaining PTM

Bio

Bio

H3

H3

Ac

tin

tin

Bi

Biotinylated
Histone H3 Ab

SA-PE

P

Intensity (MFI)Median Fluorescent

H3 pan-acetyl
8K

-2

-1

0
log[SAHA, µM]

1

2

10K
H3S10ph
8K
Intensity (MFI)

SA-PE

SA-PE

in
ot

10K

6K

Median Fluorescent

Streptavidinphycoerythrin

H3K9acSPECIFIC EFFECTS

Time-consuming Western blots can now be replaced inearly target validation programs that correlate global
histone modification enrichment or depletion withdisease. The Histone H3 PTM Multiplex Assays provide a
rapid, highly sensitive, high-throughput method for profiling
specific and off-target changes in histone PTMs in a
multiplex format. Get more data from less input, in lesstime and at a lower cost.

12K

OFF-TARGET EFFECTS

Active Motif has developed the first multiplex epigeneticas­say for studying histone post-translational modifications(PTMs). The Histone H3 PTM Multiplex Assay is designed foruse with Luminex instruments, enabling high-throughput
processing of nanogram-sized samples to simultaneouslyinterrogate multiple PTMs in a single reaction.

6K
H3K4me34K

2K

H3K27me3-2-10log[SAHA, µM]

1

2

Me3

Figure 2: The Histone H3 PTM Multiplex Assay shows increased histone acetylation in response to SAHA-mediated HDAC inhibition. HeLa cells pre-treatedwith increasing concentrations of the HDAC inhibitor SAHA were evaluatedin a multiplex of H3 pan-acetyl, H3S10ph, H3K9ac, H3K4me3, H3K27me2 &PTM Abconjugated beadH3 Total Ab-conjugated beads using the Histone H3 PTM Multiplex Assay. The
results demonstrate the ability of the assay to simultaneously assess specific andFigure1: Schematic of the Histone H3 PTM Multiplex Assay. The assay works as asolution-based sandwich ELISA to evaluate histone H3 PTM levels.

off-target effects of the treatment on histone modification levels. The dashedlines represent IC50 values determined for the acetyl marks.

ASSAY FEATURES• MULTIPLEX – the only kit available to perform
multiplexed histone modification analysis• EFFICIENT – use less input & less time than WB
to assay multiple histone PTMs
• SENSITIVE – requires only nanogram amountsof crude acid extracts or purified histones• HIGH CONTENT – simultaneously compare
specific and off-target effects in a single
reaction

To learn more about our Luminex assays, visit us at www.activemotif.com/luminex.
14

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Custom Services
epigenetic and gene regulation services
Active Motif offers a suite of services specifically designed to enhance drug development programs, especially those fo­cusedon small molecule epigenetic inhibitors. In the early phases of drug discovery our recombinant proteins and cell-based
screening services will accelerate your program, while in the late phase our genomic assays, such as ChIP-Seq, RNA-Seq,
MeDIP-Seq and Open chromatin-Seq will lead to a greater understanding of your compound’s mechanism of action.

TAKE ADVANTAGE OF OUR EXPERTISE

IDENTIFY MECHANISM OF ACTION AT THE EPIGENETIC LEVEL• Identify mechanism of action at theepigenetic level• Characterize inhibitors of epigenetic
targets• Determine the MOA and off-target
effects of your lead compounds

Figures: A) H3K27me3 Western blot shows a reduction in global H3K27me3 levels after EZH2 inhibition. B) ChIP-Seq data shows reduced H3K27me3 levels after exposure to an EZH2 inhibitor for 4 and

EPIGENETICS

8 days. The decrease in H3K27me3 signal is only revealed by using Active Motif’s ChIP-Seq spike-in
normalization protocols.

• RNA-Seq• ChIP-Seq• OPEN CHROMATIN-Seq• INTERACTOME PROFILING• HISTONE PTM QUANTITATION• DNA METHYLATION• BIOINFORMATIC SOLUTIONSGENE REGULATION• CELL-BASED PATHWAY ANALYSIS• miRNA FUNCTIONAL ANALYSIS• REPORTER ASSAY SCREENS• SEQUENCE VARIANT SERVICES

To view the full range of services and for more information, please visit us at www.activemotif.com/services.
sales@activemotif.com

15

Gene Expression Services
services for whole transcriptome sequencing and gene expression analysis
Active Motif’s transcriptome analysis services include RNA-Seq
for identification and quantitation of RNA transcripts as well
as RNA Pol II ChIP-Seq for quantitation of transcription rates toenable rapid profiling of changes in gene expression associated with transcription factor (TF) and histone modification
occupancy.

RNA-SEQ SERVICES

RNA-SEQ SERVICES
1 Prepare RNA from
cells or tissues

2 Generate cDNA

RNA-SEQ SERVICES
Simply submit RNA, cell or tissue samples. Order
RNA-Seq alone or combine with ChIP-Seq data to
uncover contextual information about:

3 Construct directional
libraries

4 Perform Next-Generation• Differential gene expression• Changes in gene structure or splicing patterns• Phenotypic effects of TF binding on gene expression

Sequencing

5 Perform data analysis

RNA-SEQ SERVICES FEATURES• PolyA selection or rRNA reduction• QC performed using Bioanalyzer• Directional library preparation

Igf1r Expression• 50bp or 100bp paired-end reads

RNA POL II ChIP-SEQ SERVICES
Analysis of RNA Pol II occupancy as a proxy measurement of transcription rates offers the advantage ofenabling you to:• Measure transcription without the influence ofRNA half-life• Identify genes poised for transcriptional activation• Generate gene expression data from cells used for
ChIP-Seq

Pol II ChIP-Seq
RNA-Seq

5

Fold Induction• Data analysis options (TOPHAT, CUFFLINKS,
CUFFCOMPARE, CUFFDIFF)

6

4
3
2
1
0

0

20

40

60

80

100

120

140

160

180 200

MinutesFigure: Gene expression profiles vary depending on the analysis method.
Data for Igf1r was extracted from RNA–Seq and RNA Pol II ChIP-Seq data
sets. Cell treatment resulted in induced gene expression that was measured
at various time points. The cumulative data show that, as measured by RNA
Pol II ChIP-Seq, transcription is induced immediately, while mRNA levels only
accumulate over time.

To view the full range of services and for more information, please visit us at www.activemotif.com/services.
16

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ChIP-Seq Services
end-to-end ChIP-Seq services
DMSO

Spike-in Normalization

Targeting epigenetic modifying enzymes for drug
development has great potential for advancing
research in the fields of cancer, inflammation,
development and neurobiology. The in-depthcharacterization of lead compounds in these areas
requires a genome-wide mechanistic perspective of
how these compounds alter epigenetic profiles.

_
Inhibitor

DMSO+

Inhibitor

Active Motif’s Epigenetic Services team specializes ingenome-wide compound characterization and offers thisassay as an end-to-end ChIP-Seq service to accelerate
your drug discovery and development program.
Figure: ChIP-Seq Spike-in Normalization Strategy reveals changes in H3K27me3
levels following treatment with EZH2 inhibitor compound. Cells treated with a

CHIP-SEQ SERVICES

small molecule inhibitor of EZH2 methyltransferase have dramatic reductions in
global H3K27me3 levels. However, H3K27me3 ChIP-Seq using standard ChIP-Seq
protocols (-) does not detect these differences. Incorporation of Active Motif’s

PROCESS STEPSChIP-Seq Spike-in Strategy (+) reveals the expected decrease in H3K27me3 ChIP-QUALITY CONTROL
1 Customer fixes cell lines

NEW: ChIP-Seq Spike-in Strategy

or freezes tissue samples

2 Active Motif prepares

Chromatin preparation isquantified and sized

chromatin and sonicates
Antibody is ChIP Qualified
prior to ChIP-Seq ChIP rxns

3 Active Motif performs
the ChIP reactions

4 Active Motif constructs
ChIP-Seq libraries
5 Libraries sequenced using
Illumina HiSeq

Seq signal.

qPCR is used to validate
success of the ChIP rxns
QC by qPCR quantificationof yield and library size
Sequencing to yield≥ 30 million tags

6 Analysis and Data Delivery

RELATED SERVICES
Active Motif also offers several other end-to-end
whole-genome services that can be combined tomeet your specific research needs.

Traditional ChIP-Seq protocols do not always detectglobal changes in histone modifications following
treatment with epigenetic inhibitors. Our novel
ChIP-Seq spike-in and normalization strategy is able
to reveal these differences and is now offered aspart of our end-to-end ChIP-Seq Service.
• Drosophila chromatin is “spiked in” to a ChIP
reaction• A second Drosophila-specific antibody is
added to reliably pull down a small fraction of
Drosophila chromatin• Drosophila tag counts are equalized acrosssamples and used to normalize ChIP-Seq data

VALIDATED ASSAYS FOR OVER 200 TARGETS• BRD4• EZH2• LSD1• H3K9ac• H3K27me3• H3K4me2• Open Chromatin-Seq – identify open regions• Low Cell ChIP-Seq – as low as 50,000 cells• RRBS – profile genome-wide DNA methylation

To view the full range of services and for more information, please visit us at www.activemotif.com/services.
sales@activemotif.com

17

Interactome Profiling
IP-mass spectrometry service for identification and characterizationof protein interactomes
The newest addition to Active Motif’s suite of custom services,
RIME (Rapid Immunoprecipitation Mass Spectrometry ofEndogenous Proteins) sheds light on the complex processof gene regulation by enabling the capture and identification of interactomes, or the associated protein networks, ofan endogenous protein of interest.

Identify interactomes regulating gene expression
Gene regulation is often oversimplified when the focus ison one particular transcription factor (TF) in any given cell
model. In reality, differential gene expression is greatly
influenced by cofactors and other protein interactions
with chromatin. RIME sorts out this complexity by providing
a means to identify the protein interactions that areimportant for gene regulation.

RIME SERVICESRIMESERVICES

1 Nuclear isolationand sonication

2 Immunoprecipitation
3 Purification of proteins
and tryptic digestion

4 Mass spectrometry

Why RIME?
Immunoprecipitation (IP) followed by mass spectrometry
has traditionally been the approach used to identify
proteins interacting with a target of interest.

5 Data analysis

RIME vs traditional IP
RIME differs from traditional IP approaches because thestarting material has been cross-linked, offering theadvantage of:1) targeting only DNA, chromatin and associated proteins,

RIME ADVANTAGES

2) enabling capture of low affinity interactions which arelost in IP/mass spectrometry,• Identify transcriptional cofactors

3) allowing more stringent wash conditions resulting in lessnon-specific interactions, and4) stabilizing protein-DNA interactions to enable thecapture of proteins that are bound at adjacent sites inthe DNA and are independent of protein-protein
interactions.

• Identify TFs bound to DNA at sites adjacent toyour target TF• Identify important targets for ChIP-Seq analysis• 50bp or 100bp paired-end reads• Detect low affinity interacting proteins

To view the full range of services and for more information, please visit us at www.activemotif.com/services.
18

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Cell-Based Pathway Analysis
identify biological responses to compound treatment
In understanding a compound’s mechanism of action it is
crucial to identify activation of both intended andunintended cellular pathways. LightSwitch™ Pathway
Screening Assays help you characterize cellular responses
to compound treatment and understand off-target effects.
Our unique collection of validated promoter reporters
measure gene expression changes associated with anumber of disease-related biological pathways. Ourstandard LightSwitch Pathway Screening Assays areavailable as stable cell lines or transfection-ready plasmids. Custom pathway sets are also available.

• Analyze 15 pathways in an HTS environment• Identify compound-mediated pathway activation• Determine the MOA and off-target effects ofyour lead compounds• Available as a product or a service
Cell Treatments
TCDD
R1881
PMA
Estradiol
DMSO
Prednisone
Heat shock
DFO
1% O2
TNF-a
IFNa
IFNg
NutlinLovastatin
Synthecol

PATHWAY PROFILING PANEL
48 promoters and controls

Tox/AhrAndrogen
CREB
Estrogen
Glucocorticoid
Heat shock
Hypoxia
NFkB
STAT
p53

Representative Reporter Constructs

Controls

log2 ratio treated/untreated

Pathway activity readout for:HIF-1aHypoxia

NFkB

Inflammation

CREB

cyclic AMP

HSF1Heat shock

p53

DNA damage, apoptosis

STATInterferon

SREBP

Cholesterol biosynthesis

ER

EstrogenAR

AndrogenGR

Glucocorticoid

AhR

Toxicity

Figure 1: LightSwitch Pathway Screening Services can profile pathway responses to treatments. The heat map above shows the inducible activity of 29
different reporter constructs that represent 10 different pathways in response to
15 different treatments.

To learn more about our LightSwitch Pathway Screening Assays and Services,
please visit us at www.activemotif.com/lightswitch.
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19

Functional Assays for miRNA Researchmeasure miRNA function and validate miRNA targets
MicroRNAs (miRNAs) are involved in many areas ofdisease biology, and are actively being tested in theclinic as drug targets and therapeutic molecules. The
LightSwitch™ 3´UTR Reporter Assay provides an HTS
system that is ideally suited for screening the effects ofsmall molecules and chemical modifications on miRNAs.

Understanding the biological role of a miRNA often
requires identification of downstream targets. The
LightSwitch 3´UTR reporter system is a validated
assay that can rapidly identify hundreds of miRNAtarget genes in a single experiment.

The LightSwitch 3´UTR Reporter GoClone™ Collection
includes over 12,000 ready-to-transfect human 3´UTRreporter vectors and 1000 synthetic miRNA target
reporters to study miRNA-3´UTR interactions, validate
miRNA targets, and measure RNA stability, translation
efficiency and the functional impact of miRNAs on agene-by-gene basis.

Figure: Interaction of miR-122 with 3´UTR targets. 3´UTR targets of miR-122 werecloned into the LightSwitch 3´UTR vector and co-transfected with either miR122 mimic or a non-targeting control miRNA into K-562 cells. The graph shows
the four strongest responders along with two non-responding controls.

• Measure the effects of small molecules or
chemical modifications on miRNA activity• Validate downstream miRNA targets• Identify all miRNAs that regulate a gene ofinterest

PRODUCTS FOR miRNA FUNCTIONAL SCREENS• 12,000 pre-cloned human 3´UTR reporters• Synthetic target reporters containing sequences
fully complementary to 1000 human miRNAs• miRNA mimics and inhibitors• miRNA screening services also available• Validated 3´UTR controls• Determine seed sequence variant effects• Complete screening service to validate 3´UTRtargets for any miRNA

For more complete information on our miRNA targets, mimics, inhibitors and services,
please visit www.activemotif.com/ls-3utr.
20

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Reporter Assays for Drug Screening
accelerate reporter assay screens with the LightSwitch™ System
The LightSwitch™ Luciferase Assay System dramaticallyreduces the time investment needed to identify themost responsive promoter to develop as a new HTS reporter assay. Over 18,000 pre-cloned human promoter
reporter vectors are available to choose from for immediate incorporation into plate-based screens.

PromoterCloned into
Reporter
Vector• 18,000 pre-cloned human promoter reporters forscreening expression changes of any human gene

400,000

RLU

300,000

RenSP
Luciferase

LightSwitch™
Promoter
Reporter• 100 transcription factor response element
reporters for screening transcription factor activity• Validated promoter reporters for measuring
pathway activation are also available (see page 21)

Gene of interest

2. Treat with test
compoundor condition.

1. TransfectVector.

EquilinEstriol
Estrone
EE2
E2

Light
Signal

200,000

3. Measureluciferase
activity.

100,000

0
-12

-11

-10

-9

-8

-7

M (log10)

Figure : Dose response of LightSwitch SYT8 Promoter Reporter construct to estrogencompounds. HT1080 cells were co-transfected with a SYT8 Promoter GoClone™
(Product ID S714388) and an ER cDNA expression plasmid, then treated with fivedifferent estrogen compounds for 24 hours before the luminescence was measured using the LightSwitch Luciferase Assay Kit.

CUSTOM SERVICES AVAILABLE• Stable cell line generation• Custom assay development• Custom cloning of novel regulatory elements(or TF response elements)

For more complete information, please visit us at www.activemotif.com/lightswitch.
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21

UNITED STATES
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Carlsbad, CA 92008 USA
Toll Free: 1 877 222 9543
Direct: 1 760 431 1263
Fax: 1 760 431 1351
sales@activemotif.com
Customer Service:orders@activemotif.com

EUROPE
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Avenue Reine Astrid, 92
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Germany Free Phone: 0800/181 99 10
France Free Phone: 0800/90 99 79
UK Free Phone: 0800/169 31 47
Other Countries, Direct: +32 (0)2 653 0001
Fax: +32 (0)2 653 0050
eurotech@activemotif.com

JAPAN
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2-21 Ageba-Cho
Shinjuku-Ku
Tokyo, 162-0824, Japan
Direct: +81 (0)3 5225 3638
Fax: +81 (0)3 5261 8733
japantech@activemotif.com

CHINA
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Bldg. 10, Suite 202
Pudong District
Shanghai, 201315, China
Hotline: 400 018 8123
Direct: +86 21 2092 6090
techchina@activemotif.com

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