Products and Services to Study:

DNA Methylation

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Products and Services to Study:

DNA Methylation
DNA methylation (5-mC) and methylation variants like 5-hmC contribute to mechanisms responsible for regulating
gene expression and many other biological processes. These epigenetic modifications have also become importantbiomarkers for cancer, aging, neurological disorders, and many other human diseases.
Active Motif is proud to be the leading provider of products and services to study DNA methylation. Our comprehensive
product line can support you whether you need just antibodies or recombinant proteins to do assays yourself or endto-end epigenetic services in which our experts perform the assays, NGS, and bioinformatics for you.

Methylated DNA Enrichment Methods
Methylated DNA Immunoprecipitation (MeDIP) and Hydroxymethylated DNA
Immunoprecipitation (hMeDIP) are techniques in which an antibody specific for either
5-mC or 5-hmC is used to selectively tag methylated genomic DNA fragments. Protein
G magnetic beads are used to capture the antibody tagged fragments. MethylCollector™
Ultra is based on the Methylated CpG Island Recovery Assay (MIRA) which utilizes a Histagged recombinant methyl-binding protein complex, MBD2b/MBD3L1, that specificallybinds methylated CpGs in genomic DNA fragments. Nickel-coated magnetic beads
capture the protein-DNA complexes. Enriched methylated DNA fragments can either be
amplified via qPCR or sequenced.
Products & Services for Genome-Wide & Gene-Specific Analysis
Product/ServiceFormat

Catalog Number

MeDIP Kit

10 rxns

55009

hMeDIP Kit

10 rxns

55010

MethylPath™ MeDIP-Seq Service

Custom-Contact Us

25025

MethylPath™ hMeDIP-Seq Service

Custom-Contact Us

25052

30 rxns

55005

MethylCollector™ Ultra Kit

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Products and Services to Study:DNA Methylation

Figure 1: MeDIP-Seq Uncovers Aging-associated Methylation Patterns in Cell-Free DNA (cfDNA).
Serum samples were collected in serum separator tubes (SST) from healthy “normal” adults, two <25 yoand two >60 yo. Cell-Free DNA (cfDNA) was purified with the Active Motif Cell-Free DNA Purification Kit(Cat no. 25503/25504) and quantified with a Qubit fluorometer. 200 ng cfDNA was used in the MeDIPKit and libraries were prepared using the Swift Biosciences Accel-NGS™ 1S PLUS DNA Library Kit (Cat no.
10024). Input to library prep was determined using AluI quantification. MeDIP and Input libraries weresequenced (38bp PE chemistry on Next-Seq500) and processed as per Active Motif’s standard pipeline
for MeDIP-Seq. Peaks called using MACS2 relative to input (p-value cutoff = 1.00x10-7).

Figure 2: NGS Data generated from MeDIP enriched DNA fragments from PBMCs.
DNA was enriched from 1 µg of adaptor-ligated human PBMC DNA using Active Motif’s MeDIP Kit. NGS
was performed using the Illumina platform to generate 26 million sequence tags. Tags were mapped togenerate a whole-genome DNA methylation profile. The image above shows that the enriched regions(purple peaks) correlate well with CpG density (tan peaks).

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Products and Services to Study:DNA Methylation

Global Analysis of DNA Methylation
Global ELISA-based DNA methylation assays are used to investigate the overall levels of 5-mC or 5-hmC
in samples to determine whether treatment conditions or disease phenotypes change epigenetic profiles.
Clinical researchers use these high-throughput assays when screening large numbers of patient samples
from their study cohorts.

Products Available to Study Global DNA Methylation Levels
ProductFormat

Catalog Number

Global DNA Methylation Assay-LINE-1

1 x 96 rxns

55017

Global 5-hmC Quantification Kit

1 x 96 rxns

55025

Figure 3: Global DNA Methylation – LINE-1 Assay shows changes in 5-mC levels in cfDNA with age.
Human serum samples from healthy “normal” adults <25 yo (N=6, mean: 21.83) and >60 yo (N=9,mean:65.89) were collected in serum separator tubes (SST). cfDNA was extracted using Active Motif’s
Cell-Free DNA (cfDNA) Purification Kit (Cat No. 25503/25504) from 5ml of serum and quantified usinga Qubit fluorometer. Following the Global DNA Methylation Line-1 Kit protocol, without MseI digestion,
25 ng of cfDNA per well was used. The % 5-mC for each sample was calculated as a percentage ofdetectable CpG residues in the LINE-1 probe region, using the methylated DNA standards included in thekit. All samples were done in triplicate. Boxplot shows 25%-75% (▢) , min-max (Ι), average (x), and median(-) % 5-mC.

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Products and Services to Study:

DNA Methylation

Single-Nucleotide Resolution Analysis of DNA Methylation
When researchers need to know precisely which cytosine residues are methylated, as well as the overalllevels of methylation, at the region(s) of interest, a DNA methylation assay with single-nucleotide resolution
is required. The gold standard methods for investigating DNA methylation at single-nucleotide resolution
involve performing bisulfite conversion.

Products & Services for Single-Nucleotide Resolution 5-mC Analysis
Product/ServiceFormat

Catalog Number

Reduced Representation Bisulfite Sequencing (RRBS)

Custom-Contact Us

25069

Targeted Bisulfite Sequencing Service

Custom-Contact Us

25035

FFPE Bisulfite Conversion Kit

40 rxns

55021

Bisulfite Conversion Kit

50 rxns

55016

Figure 4: Differential Methylation in RRBS Samples.
The displayed regions are representative regions from the genome-wide data set and shows differential
DNA methylation at an exon of CD19. Each block is a separate data point with red representing amethylated cytosine and blue representing an unmethylated base.

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Products and Services to Study:

DNA Methylation

DNA Methylation Antibodies
The best DNA methylation assays start with the best antibodies, and Active Motif offers the highest qualityand most complete line of antibodies available to study DNA methylation.

Antibodies to Study DNA Methylation
Featured Antibodies

Applications

Formats

Catalog Number

AbFlex® 5-methylcytosine antibody (rAb)

DB

100 µg91187

5-Methylcytosine (5-mC) antibody (mAb)

DB, MeDIP, MeDIP-Seq, IF

100 µg61479

5-Methylcytosine (5-mC) antibody (mAb)

MeDIP, IHC, ELISA

50 ug

39649

5-Hydroxymethylcytosine (5-hmC) antibody (mAb)

DB, MeDIP, ChIP-qPCR, ICC/IF, IHC(P)100 µg39999

5-Hydroxymethylcytosine (5-hmC) antibody (pAb)

DB, MeDIP, MeDIP-Seq, ICC/IF, IHC(P)100 µg39791

For a complete list of antibodies
visit activemotif.com/dnamethabs

DNA Methylation Enzymes
High-quality recombinant enzymes help improve development and optimization of biochemical assays,
including high-throughput drug discovery screens.

Recombinant Enzymes
EnzymeFormat

Catalog Number

Recombinant TET1 (1418-2136) protein

20 µg

31417

Recombinant DNMT1 protein

20 µg/1 mg

31404/31800

Recombinant DNMT3A protein

20 µg/1 mg

31406/31802

Recombinant DNMT3A (278-432) protein

100 µg/1 mg

31541/31941

Recombinant DNMT3A / DNMT3L complex

20 µg/1 mg

31415/31815

Recombinant DNMT3B protein

20 µg/1 mg

31413/31813

Recombinant DNMT3B (212-358) protein

100 µg/1 mg

31542/31942

Recombinant DNMT3B / DNMT3L complex

20 µg/1 mg

31416/31816

Recombinant DNMT3L protein

20 µg/1 mg

31414/31814

For a complete list of DNA Methylation Proteins & Enzymes
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Products and Services to Study:

DNA Methylation

DNA Methylation Assays
DNA methyltransferase (DNMT) enzymes are dysregulated in many cancers and other human diseases,
making them attractive drug development targets. This assay allows studying DNMT activity in a highthroughput manner.

DNA Methylation Activity Assay
ProductDNMT Activity/Inhibition Assay

FormatCatalog Number

1 x 96 rxns

55006

Fig. 5: Recombinant DNMT1 protein activity tested
using DNMT Activity/Inhibition Assay.
Assay was performed using DNMT Activity /Inhibition Assay, Active Motif Catalog No. 55006.
The substrate, AdoMet and DNMT1 were incubatedfor 1 hour at 37°C. The whole reactions weredeveloped for 4 min.

DNA Methylation Accessory Products and Controls
Successful DNA methylation assays that can be reliably interpreted require using high-quality sample
preparation methods and robust controls you can trust.

Sample Preparation & Control Products
EnzymeFormat

Catalog Number

100 ml/250 ml

25503/25504

Methylated DNA Standard Kit

3 x 2.5 µg

55008

Fully Methylated Jurkat DNA

10 µg

55003

Cell-Free DNA (cfDNA) Purification Kit

The Cell-Free DNA (cfDNA) Purification Kit isfully compatible with the KingFisher Flex andDuo. Please contact Active Motive technicalsupport for the script files.

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Products and Services to Study:

DNA Methylation
Publications
For a complete list of Active Motif Reagents and Services published for DNA Methylation Research,
visit activemotif.com/publications.

Additional Resources
Learn more by visiting our Epigenetics Resources center – a complete series of Webinars, eBooks,and TechNotes designed to help you learn more about these methods and to stay up-to-date!

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