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#49768 DNMT3A (E9P2F) Rabbit mAb

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DNMT3A 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性85, 95, 130Rabbit IgG-20℃
種交差性 (社内試験済)
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社内試験はしていませんが、配列が100%相同であるため反応すると推定される種

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49768 の推奨プロトコール i

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特異性・感度
DNMT3A (E9P2F) Rabbit mAb recognizes endogenous levels of total DNMT3A protein. This antibody detects multiple isoforms of DNMT3A, including isoform 1 and isoform 2. This antibody does not cross-react with DNMT3B or other DNMT proteins.
使用抗原
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human DNMT3A protein.

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社内データ

※下記の社内データは、すべて49768 の推奨プロトコールで実験した結果です。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using DNMT3A (E9P2F) Rabbit mAb.

IP

IP

Immunoprecipitation of DNMT3A from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DNMT3A (E9P2F) Rabbit mAb. Western blot analysis was performed using DNMT3A (E9P2F) Rabbit mAb.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either DNMT3A (E9P2F) Rabbit mAb or DNMT3A (D2H4B) Rabbit mAb #32578, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figures show binding across HUWE1, a known target gene of DNMT3A (see additional figures containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either

DNMT3A (E9P2F) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HUWE1 Intron 26 Control Primers #33770, human ARHGEF2 intron 13 primers, human SDHAP1 intron 10 primers, and SimpleChIP® Human GAPDH Exon 1 Primers #5516. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

バックグラウンド

Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2 and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor maintenance DNA methylation. DNMT1, DNMT3A and DNMT3B together form a protein complex that interacts with histone deacetylases (HDAC1, HDAC2, Sin3A), transcriptional repressor proteins (RB, TAZ-1) and heterochromatin proteins (HP1, SUV39H1), to maintain proper levels of DNA methylation and facilitate gene silencing (3-8). Improper DNA methylation contributes to diseased states such as cancer (1,2). Hypermethylation of promoter CpG islands within tumor suppressor genes correlates with gene silencing and the development of cancer. In addition, hypomethylation of bulk genomic DNA correlates with and may contribute to the onset of cancer. DNMT1, DNMT3A and DNMT3B are over-expressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast and stomach carcinomas (9-12).

使用例
 
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13451   DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific)
28692   5-Methylcytosine (5-mC) (D3S2Z) Rabbit mAb
32578   DNMT3A (D2H4B) Rabbit mAb
33770   SimpleChIP® Human HUWE1 Intron 26 Control Primers
3598   DNMT3A (D23G1) Rabbit mAb
36836   5-Carboxylcytosine (5-caC) (D7S8U) Rabbit mAb
5032   DNMT1 (D63A6) XP® Rabbit mAb
51660   5-Hydroxymethylcytosine (5-hmC) (HMC31) Mouse mAb
5516   SimpleChIP® Human GAPDH Exon 1 Primers
67259   DNMT3B (D7O7O) Rabbit mAb
7003   20X LumiGLO® Reagent and 20X Peroxide
7017   6-Tube Magnetic Separation Rack
7071   Phototope-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
74178   5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb
88989   SimpleChIP® Universal qPCR Master Mix
9002   SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
9003   SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
9004   SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads)
9005   SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)
9006   ChIP-Grade Protein G Magnetic Beads
9007   ChIP-Grade Protein G Agarose Beads
9999   Nonfat Dry Milk

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

DNMT3A (E9P2F) Rabbit mAb

Immune Cell Signaling Pathways

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