DNMT3A Polyclonal antibody

• Featured Product
• KD/KO Validated

DNMT3A Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, IP, ChIP, RIP, ELISA

Conjugate

Unconjugated

Publications

42

Cat no : 20954-1-AP

Synonyms

M.HsaIIIA, EC:2.1.1.37, EC:2.1.1.-, DNA MTase HsaIIIA, DNA (cytosine-5)-methyltransferase 3A

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Validation Data Gallery

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  WB analysis using 20954-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 20954-1-AP (DNMT3A antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 20954-1-AP

  WB result of DNMT3A antibody (20954-1-AP; 1:12000; incubated at room temperature for 1.5 hours) with sh-Control and sh-DNMT3A transfected HepG2 cells.

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  WB analysis using 20954-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 20954-1-AP (DNMT3A antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 20954-1-AP

  IP result of anti-DNMT3A (IP:20954-1-AP, 4ug; Detection:20954-1-AP 1:5000) with HeLa cells lysate 1560 ug.

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  IHC staining of human stomach cancer using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human stomach cancer using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse colon using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse colon using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse testis using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse testis using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat colon using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded rat colon tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat colon using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded rat colon tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat testis using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded rat testis tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat testis using 20954-1-AP

  Immunohistochemical analysis of paraffin-embedded rat testis tissue slide using 20954-1-AP (DNMT3A antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of K-562 using 20954-1-AP

  Immunofluorescent analysis of (4% PFA) fixed K-562 cells using DNMT3A antibody (20954-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HeLa using 20954-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using DNMT3A antibody (20954-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

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Tested Applications

• Positive WB detected in: Jurkat cells, HEK-293 cells, HepG2 cells, HeLa cells, NIH/3T3 cels, mouse testis tissue, rat testis tissue
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human stomach cancer tissue, mouse colon tissue, mouse testis tissue, rat colon tissue, rat testis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: K-562 cells, HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 4 publications below
• WB: See 36 publications below
• IHC: See 2 publications below
• IF: See 4 publications below
• ChIP: See 4 publications below
• RIP: See 1 publications below

Product Information

20954-1-AP targets DNMT3A in WB, IHC, IF/ICC, IP, ChIP, RIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, goat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: DNMT3A fusion protein Ag15111
• Full Name: DNA (cytosine-5-)-methyltransferase 3 alpha
• Calculated Molecular Weight: 912 aa, 102 kDa
• Observed Molecular Weight: 120-130 kDa
• GenBank Accession Number: BC043617
• Gene Symbol: DNMT3A
• Gene ID (NCBI): 1788
• RRID: AB_10733339
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

DNA methylation in vertebrate animals is an epigenetic modification that is important for embryonic development, imprinting, and the inactivation of X chromosomes. DNA methylation is catalyzed by a family of DNA methyltransferases (DNMTs) that include the maintenance enzyme DNMT1 and de novo methyltransferases DNMT3a and DNMT3b. The overexpression of DNMT1, DNMT3a, and DNMT3b has been reported in various malignancies, including gastric, urothelial, and lung cancers, and may be related to tumorigenesis, tumor progression, and poor survival. Two isoforms of DNMT3a exist: the full-length DNMT3a, and the shorter form DNMT3a2 which lacks the N-terminal fragment. DNMT3a is expressed ubiquitously at low levels, while DNMT3a2 is specially expressed at high levels in embryonic stem cells and shows restricted expression in tissues known to undergo de novo methylation including testis and ovary. This antibody was raised against the N-terminal region of human DNMT3a. It is expected to detect the 120-130 kDa DNMT3a but not 72-100 kDa DNMT3a2.

Publications

Species	Application	Title
human	WB,ChIP	Mol Cancer; CircNTNG1 inhibits renal cell carcinoma progression via HOXA5-mediated epigenetic silencing of Slug; Authors - Yanping Liang ; KD Validated
rat	WB	J Hazard Mater; GRP78/IRE1 and cGAS/STING pathway crosstalk through CHOP facilitates iodoacetic acid-mediated testosterone decline; Authors - Li Mou
human	IF	Sci Adv; Tumor suppressor CEBPA interacts with and inhibits DNMT3A activity.; Authors - Xiufei Chen
mouse	WB	Protein Cell; Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells.; Authors - Hongling Zhang
mouse	WB	Cell Mol Biol Lett; miR-29b-3p regulates cardiomyocytes pyroptosis in CVB3-induced myocarditis through targeting DNMT3A; Authors - Ya Wang
human	ChIP	Cell Biol Toxicol; Hypermethylation of microRNA-497-3p contributes to progression of thyroid cancer through activation of PAK1/β-catenin.; Authors - Yuxia Fan

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