TOOLS FOR DISEASE RESEARCHTARGETING EPIGENETICS & GENE REGULATION• Oncology• Inflammation• Immunology• Neuroscience

DISEASE RESEARCH TOOLBOX
antibodies, assays & services to accelerate your research

As more and more diseases are linked to epigenetic abnormalities, there has been an exponential growth in research focusedon the association between epigenetics and disease. As such, laboratories new to the areas of epigenetics and gene regulation
research are experiencing a growing need for tools to facilitate the initiation of research into these areas.
Active Motif is the industry leader in developing and delivering innovative tools to enable epigenetics and gene regulation research.
We provide the highest quality products and services to support advancements in disease research. Our staff of highly qualified
scientists work in close collaboration with world-class researchers and opinion leaders to continually develop new and innovative
tools to study chromatin, histones, DNA methylation, non-coding RNAs and transcription biology. Whether you are an expert in thefield of epigenetics or a researcher interested in integrating epigenetics research into your studies, Active Motif makes it easy tofind the right tools by providing the most comprehensive product portfolio to enable all aspects of the epigenetics workflow.

• Antibodies for Epigenetic & Disease Targets
Antibodies &Chromatin• Chromatin Immunoprecipitation Overview• ChIP for FFPE
• ChIP for Difficult-to-lyse Cells (ChIP PBMC)• Multiplex Epigenetic Assays• Global DNA Methylation Assays
Histones &DNA Methylation• Histone Modification ELISAs• DNA Methylation Analysis• Tools for Neuroepigenetic Research• Genome-wide Services• Tools for Inflammation Research• Transcription Factor Activity Assays

Transcription &miRNAs• Multiplex AP-1 and NFκB Activity Assays• Reporter Assays to Study Gene Regulation• Cell-based Pathway Screening Assays• Reporter Assays to Study miRNA
For a complete list of available products, please visit us at www.activemotif.com.

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Epigenetics & Disease
Histone H3

K12ac

K16ac

K20me3√R3me2

Multi-Kac√

Histone H4
K18ac

√

K9me1-3

K9ac

BreastK4me1-3Tumor TypeMulti-Kac

The epigenetic state of the cell is controlled by the activity ofproteins that add and remove small chemical modifications tohistones and to DNA. Aberrant epigenetic regulation can leadto changes in gene expression and human disease (Tables 1 & 2).
The link between epigenetics and disease has been substantiatedthrough the identification of mutations in, or altered expressionof, epigenetic regulator proteins that are associated with amultitude of pathological states and poor disease prognosis.

√

√

√

√

H
m ist
od on
ifi e
ca
ti
DN on
m A
et
hy
la
tio
n
N
o
RN n
c
A od
in
g

Colorectal

Learning and memory√√

Esophageal

√
√

√

√

Lung

√
√
√

√

√

√

√

√

√

Hematological
Kidney√√√Addiction√√√

Pancreatic

Autism spectrum

?

?

?

Prostate

Epilepsy√√√

Table 2: Histone modifications exhibiting altered levels in neoplastic tissue.

PTSD

√

√

Alzheimer's disease√√

Stress√√Schizophrenia√√

Bipolar disorder√√

Adult neurogenesis

√

√

√

Table 1: Examples of neural processes and disorders

These mutations have been identified in all three majorclasses of epigenetic proteins: the Writers (enzymes that
deposit modifications), the Erasers (enzymes that remove
modifications) and the Readers (proteins that recognize andbind epigenetic modifications). Recent drug development
strategies that target these enzymes have proved highly
successful, resulting in four FDA approved cancer drugs, withmany more promising leads on the horizon.
READERS
acetylases
methylases
kinases

√

√

√

√

√

√

√

associated with epigenetic modifications.

WRITERS√*

bromodomain
chromodomain
PHD finger
WD40 repeat

The epigenetic tool kit
The development of methods to perform chromatin
immunoprecipitation (ChIP) and analyses of DNA methylation,
histones and gene regulation, along with Next-Generation
sequencing tools, is allowing researchers to get a better pictureof the genome-wide changes associated with cancer and otherdiseases. Active Motif provides products that utilize familiar
techniques and reagents, or simplify technologies, to enable
even the novice to perform epigenetic studies. These include:• Antibodies to discriminate between the multiple posttranslational modifications on the histone tails and to
detect methylated DNA• ChIP and ChIP-Seq techniques to enable scientists to linkspecific states of chromatin to individual gene loci in a cell tounderstand how genes are regulated• ELISA-based methods for analysis of global changes in levels
of intercellular markers, such as histone modifications, DNA
methylation and transcription factor binding• Luciferase reporter assays to study the impact of promoter
regulation and miRNA-3´UTR interactions on thetranscriptional and post-transcriptional regulation of genes.

+

ERASERS
deacetylases
demethylases
phosphatases

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3

Antibodies
broad portfolio of disease-related targets
Active Motif develops and provides highly validated antibodies
specific for epigenetic targets and proteins involved in gene
regulation. Our antibodies have been referenced in hundredsof publications in top scientific journals. However, we are
more than just an antibody supplier. We identify, produce andrigorously test each antibody in our collection to ensure that weoffer only the highest quality product. We perform thoroughspecificity testing, including screening antibody specificity tohistones and their post-translational modifications with our
MODified™ Histone Peptide Array. We also collaborate with
top scientists that help us identify the most relevant targets andvalidate our antibodies for the most desired applications.
We monitor every step from immunogen design and specificity
screening to application validation. This thorough processensures our antibodies meet the high standards for consistency
and quality that you require to help you achieve the goals ofyour disease research.
OVER 700 DNA METHYLATION, HISTONE AND
TRANSCRIPTION FACTOR TARGETS
EPIGENETICS• H2A, H2B, H3 & H4 PTMs
• DNA methylation• Bromodomains• Methyltransferases• Demethylases• Acetyltransferases• Deacetylases

TRANSCRIPTION FACTORS• Hormone receptors• NFkB family• STAT family• IRF family• AP-1 family• p53• c-Myc• Rigorous application testing• Assay-compatible formulations• Large lot sizes• Monoclonals to histone PTMs
• Large selection of ChIP & ChIP-Seq
validated antibodies

ANTIBODIES FOR RELEVANT DISEASE AREAS
INFLAMMATIONCANCER• BRD4		
• CARM1• ER• EZH2• DOT1L• p300• AP-1• ATF-2• NFkB

NEURODEGENERATIVE• 5-hmC• APOBEC2• MeCP2• N-Myc• NRF2• TET• MAPK• NFATc1• STATIMMUNOLOGY• HDAC1• ELK• IFNA1• IRF• Nrf2• TNF

For more information and a complete list of available antibodies, please visit us at www.activemotif.com/abs.
4

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Map Disease-Specific Epigenetic Variances
simplified chromatin analysis tailored to the disease researcher
Chromatin immunoprecipitation, or ChIP, is widely used to
study changes in chromatin modifications and interactions.
Common array and whole-genome sequencing (WGS) methods
to analyze epigenetic dysregulation in human disease have anumber of shortcomings. Microarrays lack the qualitative
information offered by sequencing-based approaches. However,more comprehensive and qualitative methods like WGS areoften cost-prohibitive for disease researchers who are typicallyworking with large numbers of samples. ChIP-Seq is a highly
powerful tool in epigenetics research because it offers a

ChIP
Antibodies

Previously, ChIP analysis has been challenging for diseaseresearchers who are unfamiliar with these difficult-to-perform
techniques or who principally work with human primarytissues that present a challenge for sample preparation. At
Active Motif, we are continually expanding our portfolio of ChIP
products and services to provide innovative solutions to addressthese challenges and meet the needs of disease research.

Chromatin
IP

Downstream
Analysis

DRxn

Chromatin
Preparation

simple, genome-wide method to enable mapping of chromatinstates to specific regulatory loci.

Cycle

A wide selection oftools for sample
preparation
includinghomogenizers,
sonicators,
enzymatic shearing
kits and pre-madechromatin.

A large offering of
ChIP and ChIP-Seq
validated antibodies
for histone
modifications,chromatin
remodeling proteins
and transcription
factors (seeopposite page).

ChIP kits andaccessories for avariety of cell andtissue types anddownstream
applications (seepages 6-7).

A comprehensive
selection of qPCR
and ChIP analysis
tools and a wide
variety of Custom
Epigenetic Services
for genome-wide
ChIP and DNA
methylation
analysis (see pages
12-13).

For more information on ChIP and ChIP-related products, please visit us at www.activemotif.com/chip.
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5Uncover Epigenetic Variances in Archived Tissue
perform ChIP on FFPE primary tissue samples with ChIP-IT® FFPE
Large collections of archived
formalin-fixed, paraffin-embedded(FFPE) tissue are available thatharbor invaluable information
about the variances between
normal and diseased tissue
states. However, performing
ChIP has not been possible on FFPE samples because of the
difficulty in extracting high-quality chromatin from very limited
and degraded material. Active Motif’s ChIP-IT FFPE assay is
the first-of-its-kind to enable extraction and ChIP analysis ofchromatin from FFPE samples for Next-Generation sequencing.
Active Motif’s ChIP-IT® FFPE products
The ChIP-IT FFPE Chromatin Preparation Kit is optimized to
extract high-quality ChIP-grade chromatin from preserved
human, mouse or rat FFPE material. The kit has been successfully used to extract chromatin from FFPE blocks that werestored over 10 years in less than ideal conditions (Table 1).
For ChIP and ChIP-Seq analysis, the ChIP-IT FFPE Kit, designed
for use in conjunction with our ChIP-IT FFPE Chromatin
Preparation Kit, is the only ChIP kit available that can enrich for
ChIP DNA using FFPE-extracted chromatin. The assay produces
ChIP-enriched DNA in sufficient amounts and quality for analysis
by qPCR (Figure 2) or Next-Generation sequencing (Figure 3).

FFPE sample

Sample used per chromatin prep

Human colon

Tissue block – five 20 µm sections

Human kidney

Tissue block – twenty-five 20 µm sections

Human lung

Tissue block – two 20 µm sections

Rat whole brain

5 slides – two 5 µm sections per slide

Rat hippocampus

25 slides – two 5 µm sections per slide

Table 1: Examples of the sample material we have successfully validatedwith the ChIP-IT FFPE Chromatin Preparation method.

ChIP analysis of FFPE tissue enables insight
into the cancer epigenome
The ability to perform ChIP on FFPE samples enables validationin primary tissue of results obtained using cell lines as model
systems. In a recent study, the laboratory of Manuel Perucho*
detected DNA hypomethylation of the CpG island associatedwith the promoter of a gene (Gene X) that is upregulated incolon cancer. Figures 1-3 below show correlative bisulfite
sequencing and H3K4me3 and H3K27me3 ChIP and ChIP-Seq
data obtained with the use of the ChIP-IT FFPE Chromatin
Preparation and ChIP kits from normal and tumor samples
from two colon carcinoma patients. These results indicate that
upregulation of Gene X is associated with DNA hypomethylation
as well as increased H3K4me3 and decreased H3K27me3 levels
at the promoter region in tumors versus normal tissue.
*Alonso et al., in preparation.

400

450

500

550

600

650

700

750

800

850

900

950

1000

1050

1100

726 N
T
727 N
T

Figure 1: Bisulfite sequencing of the promoter-associated CpG islands in Gene X from normal (N) and tumor (T) samples in colon cancer cases 726 and 727.
Purple and copper circles indicate methylated and unmethylated cytosines, respectively.

H3K4me3726 Normal
726 Tumor
726 Metastasis

140
120
100
80
60
40
20
0

35

VWA2

H3K27me3
726 Normal
726 Tumor
726 Metastasis

30
25
20
15
10
5
0

Binding Events Detected / 1000 cells

160

Colon cancer case 727

VWA2

Binding Events Detected / 1000 cells

Binding Events Detected / 1000 cells

Binding Events Detected / 1000 cells

Colon cancer case 726

140
120

H3K4me3
727 Normal
727 Tumor

10080

H3K4me36040

726 Tumor

20
0

180

VWA2

Gene X

H3K27me3

TSS

727 Normal
727 Tumor

160
140
120
100
80

Figure 3: ChIP-Seq data profiles generated using ChIP-IT FFPE

60
40

ChIP’d DNA from human colon cancer case 726 show H3K4me3

20
0

VWA2

Figure 2: qPCR analysis of ChIP-IT FFPE ChIP DNA shows H3K4me3 andH3K27me3 enrichment at the Gene X locus in human colon FFPE samples.

6

H3K4me3726 Normal

enrichment at Gene X.

For more complete information on ChIP-IT FFPE products,
please visit us at www.activemotif.com/ffpe-chip.

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Evaluate Chromatin Changes in Primary Cells
ChIP-IT® PBMC for difficult-to-lyse T and B cells
Active Motif’s ChIP-IT PBMC Kit is the only ChIP kit available
to enable ChIP to be performed on difficult-to-lyse peripheral
blood mononuclear cells, or PBMCs, including lymphocytes(T cells, B cells & NK cells) and monocytes.

ChIP-IT® FFPE & PBMC ADVANTAGES• Obtain ChIP quality chromatin fromprimary cells & tissue• Sensitive enrichment of DNA fromnanogram quantities of chromatin• Optimized reagents improve signal andminimize background• Filtration based washes offer a faster,easier solution with better consistency

Because of the importance of PBMCs in cancer and immunology /infectious disease research, a robust method to extract
quality chromatin from these cells for ChIP and ChIP-Seq
analysis is highly warranted. PBMCs make it difficult to extract
quality chromatin because they are highly resistant to lysis
under conditions normally suitable for other cell types.
Active Motif scientists have developed an optimized chromatin
preparation method that yields high-quality chromatin fromprimary cells for ChIP analysis and Next-Generation sequencing.

H3K4me3

Identify epigenomic features regulating gene expression
during an immune response
Lymphocytes undergo highly dynamic changes in gene
expression programs during development and as a result of
clonal selection and expansion. ChIP-Seq analysis enables
researchers to study the molecular basis of gene regulation
during these processes by revealing the protein interactions andepigenetic modifications at regulatory loci within the genomethat are important for modulating gene expression.
Figure 1 demonstrates how ChIP-Seq data obtained using
the ChIP-IT PBMC Kit reveals a correlation between a loss in
H3K27 acetylation in the enhancer region upstream of the Cfc1
gene in mouse T cells in a mouse model harboring a deletionin a lymphoma-related gene. These results are consistent withthe characteristic repression of the T cell expression program
observed in many T cell lymphomas.

H3K4me1

H3K27ac

WT

H3K27me3

DKO

Figure 1: ChIP performed using the ChIP-IT PBMC Kit with antibodies against H3K4me3, H3K4me1,
H3K27ac and H3K27me3 histone modifications and chromatin from primary mouse T cells.
Epigenetic profiling was performed in a mouse model harboring a knockout (DKO) of a lymphoma
related gene. Within the region of the genome shown, there is a clear loss of the enhancer associated

PRODUCTS COMPATIBLE WITH ChIP-IT® FFPE & PBMC KITS
• ChIP-IT® qPCR Analysis Kit – simplifies qPCR data
analysis and enables normalization for comparison ofsamples across experiments• qPCR Primer Sets – for a multitude of gene targets
and model organisms• ChIP-Seq Antibodies – internal validation program forguaranteed success (see page 4)

H3K27ac modification at a location upstream of the Cfc1 gene (copper circle).

Product

Catalog No.

®

ChIP-IT FFPE Chromatin Preparation Kit

53030

®

ChIP-IT FFPE

53045

ChIP-IT® PBMC

53042

ChIP-IT qPCR Analysis Kit

53029

®

For more complete information on ChIP-IT FFPE products,
please visit us at www.activemotif.com/ffpe-chip.

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7

Multiplex Epigenetic Screening Assays
assay specific and off-target effects in a single well

SA-PE

in
ot

Bi

Biotinylated
Histone H3 Ab
H3Histonecontaining PTM

Ac

6000

-1

0

1

2

1

2

1

2

log[SAHA,uM]

OFF-TARGET EFFECTS
8000

H3K4me3
H3K27me2

6000

4000

2000

-2

-1

0

log[SAHA,uM]

NO EFFECT

H3Me3

PTM Abconjugated bead
Figure 1: Schematic of the Histone H3 PTM Multiplex Assay.
The assay works as a solution-based sandwich ELISA to evaluate histone H3 PTM levels.

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 amounts ofcrude acid extracts or purified histones• High content – simultaneously assay specificand off-target effects in a single reaction

8

8000

Bi

P

H3 pan-acetyl
H3K9ac

10000

in
ot

Bi

H3

SA-PE

SA-PE

in
ot

12000

-2

Median Fluorescent Intensity (MFI)

Streptavidinphycoerythrin

SPECIFIC EFFECTSMedian Fluorescent Intensity (MFI)

Profile changes in histone modifications in clinical andcompound-treated samples in multiplex
Profiling of variances in clinical or compound-treated samples
often involves large sample numbers in scarce supply. Traditional methods such as Westerns and ELISAs are limited by
low throughput, large input requirements and the inability tomultiplex the analysis of multiple targets. In contrast, the Histone
H3 PTM Multiplex Assay provides a rapid, highly sensitiveplate-based method that allows profiling of changes in histone
modification levels in multiplex. The assay’s ability to multiplex
analysis of specific and off-target effects, and its minimal cellnumber requirement of 500 - 2000 cells per microplate well,makes it ideally suited for screening and profiling of clinical orcompound-treated samples (Figure 2).

Multiplex Analysis of Specific and Off-target Effects

Median Fluorescent Intensity (MFI)

Active Motif has developed the Histone H3 PTM Multiplex
Assay, the first ever multiplex epigenetic assay that enables
interrogation of multiple post-translational modifications(PTMs) in a single reaction (Figure 1). Designed for use with
Luminex instruments, this high-throughput / high-content assay
uses only nanogram quantities of sample per well and can becompleted in 3 hours. Get more data from less input, in lesstime and at a lower cost.

12000

H3S10ph

10000

8000

6000

4000
-2

-1

0

log[SAHA,uM]
Figure 2: The Histone H3 PTM Multiplex Assay shows increased histone acetylationin response to SAHA-mediated HDAC inhibition.
HeLa cells treated with increasing concentrations of the HDAC inhibitor SAHA were
evaluated in a multiplex of H3 pan-acetyl, H3S10ph, H3K9ac, H3K4me3, H3K27me2 &H3 Total Ab-conjugated beads using the Histone H3 PTM Multiplex Assay. The results
demonstrate the ability of the assay to simultaneously assess specific and off-target
effects of the treatment on histone modification levels. The dashed lines represent
IC50 values, 4.0 μM and 4.6 μM, determined for pan-acetyl and H3K9ac, respectively.

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

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Measure Global Changes in the Epigenome
generate DNA methylation & histone profiles of your cell models
Active Motif offers a variety of high-throughput, quantitative,
simple ELISA-based assays to enable you to accurately andefficiently measure global changes in epigenetic DNA andhistone modifications in response to compounds, stimulus,
or disease-related alterations in cell signaling and regulation.

Global DNA Methylation – LINE-1

Quantify changes in global DNA methylation levels
Because changes in global methylation are a hallmark ofmany human diseases, including cancer, simpler methods
than HPLC or bisulfite sequencing are warranted for correlative
studies that analyze variances in genome-wide DNA
methylation status.
Active Motif’s Global DNA Methylation – LINE-1 Assay uses aunique hybridization approach that quantitates 5-methylcytosine (5-mC) levels at LINE-1 repeats as a surrogate measureof global methylation. This unique hybridization approach
offers better specificity and reproducibility than other available
methods that utilize non-specific passive adsorption (Figure 1).
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 screening changes inhistone H3 modification levels from purified core histones (seeProduct Ordering Information for a list of available Histone
Purification Kits) or histones isolated by acid extraction.

HISTONE MODIFICATION ELISAs FOR:
• Total Histone• H3K4me1• H3K4me2• H3K4me3• H3K9me2• H3K9me3• H3K27me1• H3K27me3• H3K9ac• H3K14ac• H3S10ph• H3S28phFigure 1: Comparative data obtained using the Global DNA Methylation – LINE 1 Assay (above) andBisulfite Sequencing (below, each clone represents one single LINE-1 transposable element ORFI
and II, accession number X52235) to analyze methylation of LINE-1 repetitive elements of normal (N,purple) and tumor (T, copper) colon samples. Data was provided courtesy of Dr. Johanna Samuelsson
and Dr. Manuel Perucho. Samuelsson et al., in preparation.

Product

Catalog No.

Global DNA Methylation - LINE-1 Kit

55017

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

Histone H3 acetyl Lys14 ELISA

53115

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

For more complete information on the Global DNA Methylation - LINE-1 Assay, visit www.activemotif.com/gdm.
To learn more about our histone analysis products, visit www.activemotif.com/hismodinfo.

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9

Profile Methylation States Associated with Disease
tools for DNA methylation analysis
DNA methylation, generated by the transfer of a methyl groupto the 5’-position of cytosine to form 5-methylcytosine, or 5-mC,
is an important epigenetic regulator of gene expression andgenomic organization. Aberrant DNA methylation is responsible
for local silencing of genes through promoter hypermethylation,a process implicated in tumorigenesis and other abnormalities,
and the ubiquitous global hypomethylation commonly observedin human cancers. There are also several methylation
variants thought to have their own unique epigenetic functions.
One of these, 5-hmC, is highly expressed in the brain during
development and is thought to have a unique biological rolein the Central Nervous System. However, the role of this andother methyl variants is still not fully understood.
The development of methods such as DNA methylationenrichment techniques and bisulfite conversion, along with
Next-Generation sequencing tools, is allowing researchers to
get a better picture of the genome-wide and gene-targetedchanges that occur in DNA methylation and its association withdisease. Active Motif offers a number of tools and services tosimplify and streamline DNA methylation analysis.

Antibodies &Enzymes

Bisulfite
Conversion

Methylated DNA
Enrichment

Quantitative
Assays

A comprehensive
portfolio of
antibodies (seepage 4) and proteinsfor 5-mC and
DNA Methylation
variant (5-hmC,
5-fC, 5-caC & 3-mC)
analysis.

Bisulfite Conversion
Kits and Sequencing
Services to enable
analysis ofmethylation
patterns todifferentiate
between methylated
and unmethylatedsequences (seeopposite page).

A broad selection of
kits for enrichmentof methylated DNA
variants (5-methylcytosine, or 5-mC
and 5-hydroxymethylcytosine, or
5-hmC) (seeopposite page).

Simple, highthroughput ELISAbased assays formeasuring changes
in global DNA
methylation levels(see page 9) or
DNMT activity.

For more information on DNA methylation-related products, please visit us at www.activemotif.com/dna-methylation.
10

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Tools for Neuroepigenetic Research
products to study DNA methylcytosine variants
5-Hydroxymethylcytosine (5-hmC) is highly abundant in the
CNS where it regulates various processes, including development,
aging and neuroplasticity (Figure 1). High 5-hmC levels in thedeveloping brain are associated with maintenance of active
transcription. In adult neurons, alternating DNA methylationand demethylation are observed in response to neuronal ac­tivity
suggesting 5-hmC is involved in cognitive function and psychiatricdisorders such as stress-induced PTSD. Also, aberrant 5-hmC
distribution is observed in various neurodegenerative disorders,
including Alzheimer’s and Huntington’s disease.

Profile changes in 5-hmC status at specific loci
Active Motif offers two methods for hydroxymethylated DNA
enrichment for downstream 5-hmC analysis by PCR or
sequencing. Both utilize magnetic beads for faster processing.

Gene Ontology of Top 2500 5-hMeDIP Genes

Figure 1: Gene Ontology analysis of the 2,500 genes selected with the highest hMeDIP signals.
The analysis shows that the top selected genes were biased toward brain expression. In addition,
the other most statistically significant observed ontologies were also subsets of the brain. These• hMeDIP Kit – an antibody-based technique that utilizes a
5-hmC antibody to immunoprecipitate DNA fragments containing 5-hmC from genomic DNA. The assay works with both
single-stranded and double-stranded DNA (Figures 1 and 2).

results show that 5-hmc enrichment correlates well with expected gene expression.

• Hydroxymethyl Collector™ Kits – utilize a β-Glucosyltransferase enzyme to label 5-hmC residues within doublestranded DNA fragments with a modified glucose moiety toenable distinction from other cytosine modifications during
enrichment. This sensitive technique can enrich DNA fragments containing as little as two 5-hmC residues.
• Methylation variant profiling services – Active Motif’s
Custom Epigenetic Services include genome-wide hydroxymethylcytosine (hmC), formylcytosine (fC) and carboxylcytosine (caC) profiling. For more complete information,
go to page 13 or visit www.activemotif.com/meth-variant-svc.
Differentiate between methylated & unmethylated sequences
Active Motif has a selection of kits for bisulfite conversion
analysis and determination of DNA methylation status:• Bisulfite Conversion Kit – provides single nucleotide
resolution information of the methylation status of yourenriched DNA. See page 13 for related custom services.
• ChIP-Bis-Seq Kit – takes it one step further by combining
bisulfite conversion with ChIP to enable you to obtain
DNA methylation profiles focused on your specific genomic
region(s) of interest.

Figure 2: hMeDIP-Seq performed on human brain.
hMeDIP was performed with 5-Hydroxymethylcytosine (5-hmC) antibody (Catalog No. 39999)
and human brain DNA to identify regions enriched for 5-hmC. The image above shows an 8 Mb
region on chromosome 5 with 5-hmC enrichment across several gene bodies.

Product

Catalog No.

5-Hydroxymethylcytosine (5-hmC) antibody

39769

5-Methylcytosine (5-mC) antibody

61225

hMeDIP Kit

55010

MeDIP Kit

55009

Hydroxymethyl Collector™

55013

Hydroxymethyl Collector™-Seq

55019

MethylCollector™ Ultra

55005

HypoMethylCollector™

55004

Bisulfite Conversion Kit

55016

ChIP-Bis-Seq Kit

53108

To learn more about products for 5-hmC analysis, please visit us at www.activemotif.com/hmc.
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11Custom ChIP and ChIP-Seq Services
let the ChIP experts do the work for you
Whole genome analysis using Next-Generation sequencing
has significantly broadened the ability of researchers tounderstand how epigenetic events influence disease.
However, the technical and bioinformatics challenges
associated with generating whole-genome data may make
ChIP-Seq beyond your reach.
Active Motif’s Epigenetic Services team provides a wide varietyof ChIP-Seq Services to make it possible for you to utilize ourexpertise and research tools without having to be an expert
in the techniques yourself. As the only end-to-end ChIP-Seq
provider, we have performed services using state-of-the-art
techniques for over 10 years to help accelerate the researchof scientists in academic laboratories, government institutions,biotechnology and pharmaceutical companies.

Map genome-wide chromatin interactions and modifications
associated with disease
ChIP-Seq services enable you to perform whole genome analysisof chromatin interactions and/or modifications influencing
disease propensity, outcome or responsiveness to therapies.
Figure 1 shows ChIP-Seq data revealing details about SRC3
genomic interactions that potentially influence its ability tocause resistance to hormone therapy in breast cancer.
OBTAIN ChIP-SEQ EPIGENETIC PROFILES OF:• Epigenetic inhibitor treated cells• Primary human tumors• Xenograft models

Control

ACTIVE MOTIF EPIGENETIC SERVICES

SRC3
ChIP-Seq
Estrogen• Experience – thousands of genome-wide data sets generated• Quality – QC steps ensure high-quality data
• Support – all services include bioinformatics analysis

Control

Pol IIChIP-Seq
EstrogenRET
q11.2142,880,000

ChIP-Seq Services

42,900,000

42,920,000

42,940,000

Figure 1: ChIP-Seq analysis of breast cancer cells reveals link between SRC3 activity and resistance toendocrine therapy. ChIP-Seq was performed using chromatin from control and estrogen-treated MCF-7

PROCESS STEPS

cells using antibodies against RNA pol II and SRC3, a protein that is upregulated in breast cancer and

QUALITY CONTROL

associated with resistance to endocrine therapy. The estrogen-induced SRC3 binding observed at thepromoter and gene body of the RET gene (top panel) correlates with induced transcription of the RET

1 Customer fixes cell lines
or freezes tissue samples.

2 Active Motif prepareschromatin and sonicates.

gene as measured by RNA pol II occupancy (bottom panel). RET is overexpressed in some ER-positive
Chromatin preparation isquantified and sized.

breast cancers and is known to influence the ER-mediated response to endocrine therapy.

Antibody is ChIP Qualified
prior to ChIP-Seq ChIP rxns.
3 Active Motif performs
the ChIP reactions.

qPCR is used to validate
success of the ChIP rxns.

4 Active Motif constructs
ChIP-Seq libraries.

QC of yield and library size.

5 Libraries sequenced on
Illumina platform.

Sequencing to yield≥ 30 million tags.

ACTIVE MOTIF ChIP-SEQ SERVICES• FactorPath™ ChIP-Seq – discover, identify andquantitate transcription factor and cofactor binding sites• HistonePath™ ChIP-Seq – map histone modifications
or histone modifying enzymes across the genome• TranscriptionPath™ ChIP-Seq – measure transcription
rates globally as a function of RNA pol II occupancy

6 Analysis and Data Delivery.

12

www.activemotif.com

End-to-end DNA methylation Services
send us your samples, we send you the data

• MeDIP-Seq – enrichment of methylated DNA with ahighly specific 5-methylcytosine antibody• MethylCollector™ Ultra-Seq – based on the patented
MIRA (Methylated CpG Island Recovery Assay) technology• hMeDIP-Seq – enrichment using the most specific,
highly cited 5-hydroxymethylcytosine antibody• fC & caC Genome-wide Profiling – enrichment using a
5-formylcytosine or a 5-carboxylcytosine antibody,followed by Next-Generation sequencing• Bisulfite Sequencing – detects the methylation statusof select genomic regions at single-base-pair resolution

Profile DNA methylation variances to identify biomarkers andgain insight into disease and drug mechanisms
Active Motif offers a comprehensive selection of end-to-end
DNA Methylation Services, including methylated DNA
enrichment, methyl variant profiling and bisulfite sequencing,for genome-wide DNA methylation analysis.
DNA methylation occurs mainly at CpG sites. CpG islands are highCpG density regions commonly found within the promoters ofmammalian genes. Most variance in DNA methylation observed
between tissue types and in normal versus diseased tissue occurs
a short distance from the CpG islands at “CpG shores” (Figure 1).
Profiling variances in these regions at a genome-wide scale iscrucial to understanding disease epidemiology and to identifying
potential targets for therapeutic intervention. Our Epigenetic
Services group makes it easy by doing the work for you fromsample preparation all the way to bioinformatics analysis of data.
CpG density

MethylCollector peaks• Targeted Next-Generation Bisulfite Sequencing- Multiplex many samples and multiple amplicons

into a single Next-Generation sequencing reaction- 100X to 10,000X coverage of each amplicon- No cloning bias
- Multiplexing makes large experiments cost-effective
NormalPatients72

1

2

DiseasedPatients
3

4

5

6

Disease Model
Cell Lines7

8

9

Figure 2: Targeted Next-Generation
Bisulfite Sequencing was performed48

using 9 separate samples and
CpG Site

ACTIVE MOTIF DNA METHYLATION SERVICES

Bisulfite Sequencing Services
Bisulfite Sequencing is the only method that enables detectionof the methylation status of individual cytosines at singlebase-pair resolution. Active Motif Epigenetic Services offers
two Bisulfite Sequencing options:

10 different primer pairs.
All 90 amplicons were sequenced ina single Next-Generation sequencing

24

run. This heat map shows methylation
data from one of the ten primer pairs
across the population of 9 samples.

1
0

50

% DNA Methylation

100• Sanger Bisulfite Sequencing
This traditional bisulfite approach requires cloning of bisulfite
converted amplicons and sequencing of 8 to 16 clones.

- Service includes all steps from primer design to analysis- Recommended for small-scale experiments of < 5 samples
CpG island
Cell
Type 1Figure 1: MethylCollector™ Ultra-Seq data shows DNA methylation at CpG shores in PBMC
enriched methylated DNA. Next-Generation sequencing was performed on DNA enriched from human
PBMC DNA using MethylCollector™ Ultra, and tags were mapped to generate a whole-genome DNA methylation profile. Data show DNA methylation is detected at CpG shores rather than in the CpG island itself.

CellType 2

1
2
3
4
5
6
7
1
2
3
4
5
6
7

Figure 3: Bisulfite sequencing reveals
differential methylation in two cell lines.
Sanger bisulfite sequencing was performed
on customer-selected genomic locations.
This data compares the methylation stateof a region with 24 CpGs in two different
cell lines. Seven clones from each of thecell lines were sequenced.

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

13

Tools for Inflammation Research
products to study immune function and inflammatory response
The inflammation / immune response is a highly orchestrated
process that involves multiple cellular signaling pathways
regulated by the NFκB, AP-1 and STAT transcription factor
families, which in turn are regulated by numerous cytokine
proteins including TNF-a and interferons. In a healthy human,
it protects its host from foreign invasion, toxins, and tumor
formation. However, unregulated inflammation and aberrant
immune function can damage normal cells and tissues andcan cause the onset of human disease, including autoimmune
disorders and cancer.
Active Motif offers a large number of products and services
that can help you study the regulation and function of
inflammation proteins.
Measure activity of NFκB, STAT and other transcription
factors regulating inflammation
Active Motif offers two methods for quantitative measurement oftranscription factor-DNA binding activity of immunoresponsive
transcription factors.

Figure 1: The canonical and non-canonical NFkB cell signaling pathways.

• TransAM® Transcription Factor Assays – a non-radioactive
plate-based method that measures transcription factorDNA binding activity in mammalian tissue and cell extracts.
Assays are available for over 40 different targets including
NFκB, STAT and MAPK (see opposite page).
• Transcription Factor Multiplex Assays – utilizes Luminex®
xMAP® technology to enable multiplexing for comparative
analysis of DNA binding between proteins of the same family.
Kits are available for the study of AP-1 and NFκB transcription
factor families (page 16).
Tools to study transcriptional and post-transcriptional
regulation of immunoresponsive genes
Active Motif’s LightSwitch™ Luciferase Assay System is acomplete solution for performing gene regulation studies inliving mammalian cells.
• LightSwitch™ Pathway Screening Assays – cell-based
screening assays to measure the effects of compounds ortreatments on the expression of genes associated with anumber of disease-related biological pathways (see page 18).
• LightSwitch™ 3´UTR & miRNA Collection™ – pre-cloned
luciferase 3´UTR reporter constructs and miRNA mimics &inhibitors for performing miRNA target validation studies ofinflammation genes (Table 1 & page 19).

14

miRNA

Target(s)FunctionmiR-125b

TNFα

Host immunity

miR146a

TRAF6, IRAK

Suppression of IL and
TNFα expression

miR-142-3p

N-WASP, IRAK-1

Phagocytosis, TLR signaling

miR-128a

TGFb

TGFb signaling, cell survival

miR-99b

TNFαBacterial burden

miR-155

SHIP1

Enhanced TNFα activity

miR-466l

IL-10

Phagocytosis, TLR signaling

miR-124a

CDK2, MCP1

Proliferation, monocyte
chemoattractant secretion

Table 1: miRNA protein targets and function in regulation of inflammation.

To learn more about our NFκB-related antibodies & assays,
visit us at www.activemotif.com/nfkbtools.

www.activemotif.com

Measure Transcription Factor Activity
quantitative ELISAs for transcription factor activation
TRANSAM® TRANSCRIPTION FACTOR ASSAYS
Active Motif’s TransAM Transcription Factor Assays
enable you to assess changes in transcription factor-DNA
binding activity for over 40 different transcription factors
important for inflammation, cancer, development, stem
cell biology and other disease research areas.

TransAM ASSAY FLOW CHART

Add nuclear
extract

Add primary
antibody

Add anti-IgG
HRP conjugate

1 hr.

1 hr.

Add developing
and stop solution

1 hr.

Assays are available for:• c-Myc• NFkB• IRF-3• HIF-1• STAT• AP-1• PPARg• MEF2• ER• ATF-2• AP-1• p53• Nrf2• MAPK• Oct4• and more...

Cell extractcontaining activatedtranscription factor

Oligonucleotide
coated plate(Flexi Kits enable any
oligo to be coated)

Figure 1: Flow chart of the TransAM process.
Activated transcription factor in the cell extract binds to the consensus-binding site on the oligo
immobilized in the well. Incubation with the supplied primary and secondary antibodies
specifically quantifies the amount of activated transcription factor.

TransAM Transcription Factor Assays are highly sensitive
non-radioactive assays that facilitate the study of transcription
factor-DNA binding activity in mammalian tissue and cell
extracts. Small changes in transcription factor activity levels
can have a significant impact on cellular function. Therefore, it
is important to use a sensitive assay to measure these changes.
TransAM Kits combine a fast, user-friendly high-throughput
assay for measuring transcription-factor DNA binding activity
with supreme sensitivity and specificity (Figure 1). TransAM
assays are up to 100-fold more sensitive than gelshift
assays and also yield more quantitative results (Figure 2).
Come see why we are the most cited kit on the market withover 900 citations, and counting.

TransAM ADVANTAGES:• Up to 100-fold greater sensitivity thangelshift assays• Results in less than 5 hours• Non-radioactive, colorimetric readout thatis easily quantified by spectrophotometry• No cloning or cell transfections required• Ability to assay tissue samples• 96-stripwell format compatible withhigh-throughput automation

Figure 2: TransAM Kits are more sensitive than gelshift.
Human fibroblast WI-38 cells are stimulated with IL-1a for 30 minutes. Increasing amounts ofcell extract are assayed using the TransAM NFkB p50 Kit (A) or gel retardation (B). The TransAM
method is 10-fold more sensitive and provides more quantitative data than gelshift.

For more information and an up-to-date list of available TransAM kits, please visit us at www.activemotif.com/transam.
sales@activemotif.com

15

Multiplex AP-1 & NFkB Assays
study activity of transcription factors regulating inflammation
Active Motif partnered with Luminex®, the industry leader for
multiplexing, to develop Transcription Factor Multiplex Assays
for studies of AP-1 and NFκB families.

Traditional methods to assay transcription factor activity, suchas Western blot and ELISA, are limited by the inability to quantify DNA binding events or to simultaneously compare activityof members of the same transcription factor family.

Median Fluorescent Intensity (MFI)

Simultaneously identify responsive and non-responsive AP-1
and NFκB transcription factor targets
The AP-1 family of transcription factors plays a large role in
the regulation of differentiation, proliferation and apoptosis, inparticular in relation to inflammation and cancer. Likewise, the
NFκB family of transcription factors is one of the most widely
studied protein families in immunology and cancer research.
However, comparative analysis of the relative activities or
responses of members within the same transcription factor
family have been encumbered by the lack of multiplexing
capability of the currently available technologies to study them.

3000

2500

Blank
Raji
Raji + wild-type oligo
Raji + mutant oligo

2000

1500

1000

500

0
p65

p50

RelB

p52

NFκB antibody-conjugated beads

Figure 2: Multiplex analysis of NFkB DNA binding activity using the Transcription
Factor Multiplex Kit – NFkB.
The graph shows a multiplex assay in which antibody-conjugated beads for
multiple analytes were combined into a single well for simultaneous comparison

Active Motif’s Transcription Factor Multiplex Assays overcome
these limitations. These high-throughput / high-content assays
are designed as pull-down reactions utilizing the revolutionary
Luminex xMAP® technology to enable you to quantitatively
measure the DNA binding activity of multiple AP-1 or NFκB
family members in multiplex (Figures 1 and 2). The plate-based
design makes it ideally suited for high-throughput large cohort
studies and compound screens and enables you to simultaneously assess for responsive / non-responsive or functional /non-functional AP-1 and NFκB transcription factor targets in asingle well.
Streptavidin-phycoerythrin

SA-PE

in
ot

Bi

Biotinylated TF consensus
binding sequence

Transcription factor (TF)
TF antibody-conjugated beadof the activity of various NFkB proteins within the same sample.

Product

Catalog No.

Transcription Factor Multiplex Kit – AP-1

33100

c-Fos-conjugated beads

33101

FosB-conjugated beads

33102

JunB-conjugated beads

33104

JunD-conjugated beads

33105

Transcription Factor Multiplex Kit – NFkB

33110

RelB-conjugated beads

33111

NFkB p52-conjugated beads

33112

NFkB p65-conjugated beads

33113

NFkB p50-conjugated beads

33114

Figure 1: Bead-based Transcription Factor Multiplex Assay.
In the bead-based pull-down assay, lysates are combined with a biotinylated consensus
binding sequence for the transcription factor (TF) family of interest. Fluorescent-labeledmagnetic beads conjugated to antibodies specific for each TF, each with a unique signal,
are added to capture the analyte(s) of interest. Streptavidin-phycoerythrin (SA-PE) isthen added to bind biotin. Fluorescent signals from the beads and SA-PE are used tomeasure the levels of transcription factor-bound DNA with a Luminex instrument.

16

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

www.activemotif.com

Functional Validation of Molecular Interactions
reporter assays to study gene regulation
The LightSwitch™ Luciferase Assay System provides a quick
and efficient method to measure the functional response ofpromoters to stimulus in nearly any cell-based system. The
LightSwitch promoter reporter GoClone™ collection includes
transfection-ready luciferase reporter constructs utilizing
the novel RenSP luciferase technology for over 18,000 humanpromoters and highly optimized LightSwitch reagents.
• 18,000 pre-cloned human promoter reporters forscreening expression changes of any human gene• 100 transcription factor response element
reporters for screening transcription factor activity• Validated promoter reporters for measuring
pathway activation are also available (see page 18)

PROMOTER

GENE OF INTEREST

Pre-cloned intoa Reporter
VectorRenSP
Luciferase

LightSwitch™
Promoter Reporter

1. Transfect vector.

O
N

N

N

Measure induction of select promoters in response totreatments or compounds
Much of disease research is focused on binding events,
whether they are transcription factor studies, protein-protein
and protein-DNA interactions, or analysis of the formation
or disruption of these molecular interactions under various
conditions or treatments. Active Motif’s GoClone collections oftransfection-ready human promoters, 3´UTRs and enhancer
elements along with our LightSwitch luciferase Assay Systemprovide a highly robust method to analyze the functional consequences of these interactions on a gene-by-gene basis (Figure 1).

400,000

RLU

300,000

EquilinEstriol
Estrone
EE2
E2

O
N

S
Cl

2. Treat withcompound.

3. Measure luciferase activity to determine

200,000

functional effects.

100,000

0-12

CUSTOM SERVICES AVAILABLE
-11

-10

-9

-8

-7

M (log10)
Figure 1: Dose response of LightSwitch SYT8 Promoter Reporter construct toestrogen compounds. HT1080 cells were co-transfected with a SYT8 Promoter• Stable cell line generation• Custom assay development• Custom cloning of novel regulatory elements(or TF response elements)

GoClone™ (Product ID S714388) and an ER cDNA expression plasmid, then treatedwith five different estrogen compounds for 24 hours before the luminescence wasmeasured using the LightSwitch Luciferase Assay Kit.

For more on the LightSwitch Luciferase Assay Systemand products, visit us at www.activemotif.com/lightswitch.
sales@activemotif.com

17

Identify Biological Responses to Compound Treatments
cell-based pathway screening assays• Analyze 15 pathways in an HTS environment• Identify compound-mediated pathway activation• Determine the mechanism of action (MOA) and
off-target effects of your lead compounds• Available as a product or a service• Secondary screening, “Hits-to-Lead”• Off-target analysis• Lead compound validation / optimization• Dose-response analysis

Cell Treatments

Controls
Tox/AhrAndrogen
CREB
Estrogen
Glucocorticoid
Heat shock
Hypoxia
NFkB

DISEASE PROFILING PANEL

STAT

48 promoters and controls

p53

Representative Reporter Constructs

Measure the effects of your compounds on a combination of
biologically relevant pathways
Our in-house experts use our unique collection of validated
assays to measure gene expression changes associated with anumber of disease-related biological pathways. Leverage ourunique collection of validated promoter reporter vectors fromvarious pathways: Tox/AhR, Androgen, CREB, Estrogen,Glucocorticoid, Heat Shock, Hypoxia, NFkB, STAT, SREBP, andp53 (Figure 1). Our standard LightSwitch Pathway Screening
Assays are available as stable cell lines or transfection-readyplasmids. Custom pathway sets are also available.

A SOLUTION FOR:

TCDD
R1881
PMA
Estradiol
DMSO
Prednisone
Heat shock
DFO
1% O2
TNF-a
IFNa
IFNg
NutlinLovastatin
Synthecol

To understand a compound’s mechanism of action it is crucial
to identify activation of both intended and unintended cellularpathways. LightSwitch™ Pathway Screening Assays help you
characterize cellular responses to compound treatment andunderstand off-target effects.

log2 ratio treated/untreated

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

18

HIF-1aHypoxia

NFkB

Inflammation

CREB

cyclic AMP

HSF1Heat shock

p53

DNA damage, apoptosis

STATInterferon

SREBP

Cholesterol biosynthesis

ER

EstrogenAR

AndrogenGR

Glucocorticoid

AhR

Toxicity

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

Measure miRNA Function and Validate miRNA Targets
functional reporter assays for miRNAs
MicroRNAs (miRNAs) are important regulators of gene
expression and have been shown to play a role in numerous
biological processes such as cellular signaling, development,
and apoptosis. Mutations and improper regulation of miRNAs
have been linked to a variety of physiological disorders such as
cancer and heart disease. The LightSwitch™ 3´UTR Reporter
Assay provides an HTS system that is ideally suited for screening
the effects of compound treatments or other modifications on
miRNA function.

• Measure the effects of small molecules or
chemical modifications on miRNA activity• Validate hundreds of miRNA targets in a single
experiment• Identify all miRNAs that regulate a gene of interest• miRNA screening services also available
Predicted 3´UTR targets

log2 ratio of targeting/non-targeting
miRNA luciferase activity

1.5
1.0
0.5
0.0
-0.5
-1.0

Positive control 2-1.5

Positive control 1

-2.0
-2.5

Figure 1: Results from functional screen of predicted miR-122 targets in HT-1080 cells.
The log2 ratio observed for each tested UTR reporter in the presence of the miR-122
mimic over the luminescence when transfected with the non-targeting control shows
58/142 (40.8%) of the predicted targets were significantly altered in the mimic

Measure RNA stability, translation efficiency and thefunctional impact of miRNAs on a gene-by-gene basis
Genome-wide transcript analysis can identify candidate target
transcripts but cannot measure both the changes in atranscript’s stability or translational efficiency attributable tomiRNAs. With the LightSwitch 3´UTR Reporter GoClone™
Collection and Assay System, we have created a genome-widelibrary of human 3’UTR-luciferase reporter constructs to enable
researchers to screen thousands of potential miRNA targets
in high-throughput. Figures 1-3 demonstrate how, using thisstrategy, we were able to identify new targets of miR-122, an
important regulator of cholesterol and fatty-acid metabolism
in liver that has been implicated as a therapeutic target for
metabolic disease.

co-transfection (P<0.05, t-test). Furthermore, of those 3’UTRs with significantly alteredluminescence, 25/58 (43.1%) were repressed 2-fold or more by the miR-122 mimic.

Figure 2: Knock-down of 3’UTR
Relative luciferase activity

luciferase changes in a dose-dependent
manner. Highly repressed targets fromthe above screen were tested at miRNAconcentrations ranging from 0.0625 to
100 nM. The tested 3’UTR reporters
responded to miR-122 mimic in adose-dependent fashion, with EC50 ator below 1.5 nM.

miR-122 concentration (nM)0

PRODUCTS FOR miRNA FUNCTIONAL SCREENS

3´UTR targets. 3´UTR targets of
miR-122 were cloned into theLog2 Ratio (miRNA/control)• 12,000 pre-cloned human 3´UTR reporters• Synthetic target reporters containing sequences fullycomplementary to 1000 human miRNAs• miRNA mimics and inhibitors• Validated 3´UTR controls• Complete screening service to validate 3´UTR targets
for any miRNA

Figure 3: Interaction of miR-122 with

-0.5

-1.0

LightSwitch 3´UTR vector and cotransfected with either miR-122 mimic-1.5

or a non-targeting control miRNA in
K-562 cells. The graph shows the four

-2.0

strongest responders along with two-2.5

non-responding controls.

-3.0

-3.5
o

N
rt
se

In

1

3C

D1

3

KR

IA

PP

AN

PC
G6

A2
PD

GN

S1

M

RI

To learn more about available miRNA targets, mimics, inhibitorsand services, visit us at www.activemotif.com/ls-3utr.
sales@activemotif.com

19

Enabling Epigenetics Research

NORTH AMERICAJAPAN

EUROPE

CHINA

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Direct: 760 431 1263
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