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#44145 DNMT3B (E9X7R) Rabbit mAb (Mouse Specific)

 

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2019年1月21日15時15分 現在
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DNMT3B 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性105Rabbit IgG-20℃
種交差性 (社内試験済)
交差する可能性がある種 i

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

マウス -
44145 の推奨プロトコール i

最適な結果を得るために:Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。
推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。

注:各製品に最適化されたプロトコールをリンクしています。

 

ウェスタンブロッティング (1:1000)免疫沈降 (1:50)免疫蛍光細胞染色 (IF-IC) (1:1600)クロマチン免疫沈降 (1:50)

CST推奨プロトコールに欠かせない関連製品

特異性・感度
DNMT3B (E9X7R) Rabbit mAb (Mouse Specific) recognizes endogenous levels of total DNMT3B protein. This antibody does not cross-react with DNMT3A or DNMT1.
使用抗原
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Phe324 of mouse DNMT3B protein.

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社内データ

※下記の社内データは、すべて44145 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 mES, F9, and NIH/3T3 cell lines using DNMT3B (E9X7R) Rabbit mab (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, NIH/3T3 cells show very low to no expression of DNMT3B.

IP

IP

Immunoprecipitation of DNMT3B from F9 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DNMT3B (E9X7R) Rabbit mAb. Western blot analysis was performed using DNMT3B (E9X7R) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of mES cells (left, positive) and NIH/3T3 cells (right, negative) using DNMT3B (E9X7R) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Cells were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from mES cells and either DNMT3B (E9X7R) Rabbit mAb (Mouse Specific) 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® Mouse Exo1 Exon 12 Primers #49170, mouse FH1 intron 9 primers, mouse RCC2 exon 3 primers and SimpleChIP® Mouse Oct-4 Promoter Primers #4653. 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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36836   5-Carboxylcytosine (5-caC) (D7S8U) Rabbit mAb
4653   SimpleChIP® Mouse Oct-4 Promoter Primers
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7003   20X LumiGLO® Reagent and 20X Peroxide
7017   6-Tube Magnetic Separation Rack
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7074   Anti-rabbit IgG, HRP-linked Antibody
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76853   SimpleDIP™ Methylated DNA IP (MeDIP) Kit
7727   Biotinylated Protein Ladder Detection Pack
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
95176   SimpleDIP™ Hydroxymethylated DNA IP (hMeDIP) Kit
9999   Nonfat Dry Milk

Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
ProLong is a registered trademark of Life Technologies Corporation.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

本製品は試験研究用です。

DNMT3B (E9X7R) Rabbit mAb (Mouse Specific)

Metabolic Reprogramming in Disease

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