Invitrogen

DNMT3B Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

6 Published Figures

10 References

View all (20) DNMT3B antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

DNMT3B Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of DNA Methyltransferase 3 showing positive staining in the nucleus and cytoplasm of paraffin-treated Human breast carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a DNA Methyltransferase 3 polyclonal antibody (Product # PA1-884) diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

DNMT3B Antibody in Western Blot (WB) — Western blot analysis of DNA Methyltransferase 3 was performed by loading 25 µg of Hela cell lysates and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4°C overnight. The membrane was probed with a DNA Methyltransferase 3 polyclonal antibody (Product # PA1-884) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Results show a band at 97 kDa.

DNMT3B Antibody — DNMT3B is required for de novo DNA methylation of IAPs and gene promoters upon LSH expression in Lsh−/− MEFs. (A) Expression of DNMT3B in Lsh−/− MEFs expressing either a non-silencing shRNA (shNS) or shRNAs (sh1, sh2 and sh3) targeting DNMT3B as assessed by quantitative RT-PCR. The error bars denote standard deviation. (B) Western blots showing the presence of 3xFLAG-tagged wild-type LSH in Lsh−/− clonal MEF lines expressing either non-silencing shRNA (shNS) or shRNA targeting DNMT3B (sh2 D3B). HDAC1 and MRE11 were used as loading controls. (C) Bisulfite DNA sequencing of IAPs and the promoters of Rhox2a and Rhox6 genes in indicated cell lines. Methylated CpGs are shown in black. (D) Expression relative to Gapdh of IAPs, Rhox2a and Rhox6 in indicated cell lines. The expression detected in Lsh−/− MEFs was designated as 1. The error bars denote standard deviation.

Product Details

PA1-884

Applications

Tested Dilution

Publications

Western Blot (WB)

2.0 µg/mL

View 10 publications 10 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1,000

Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to the residues M(1) K G D S R H L N E E E G A(14) C of mouse DNMT3b

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2277463

Product Specific Information

PA1-884 detects DNA methyltransferase 3 from human and mouse tissues and cells as well as recombinant human Dnmt3. This antibody has been used to detect both recombinant and endogenous DNMT3b and recombinant DNMT3a.

PA1-884 has been successfully used in Immunohistochemistry (paraffin) and Western blot procedures. By Western blot, this antibody detects DNMT3 from P19 nuclear extracts.

The PA1-884 immunogen is a synthetic peptide corresponding to residues 1-14 of mouse DNMT3b. This sequence is 79% conserved between human and mouse. PA1-884 immunizing peptide (Cat. # PEP-117) is available for use in neutralization and control experiments.

Target Information

CpG methylation is an epigenetic modification that is important for embryonic development, imprinting, and X-chromosome inactivation. Studies in mice have demonstrated that DNA methylation is required for mammalian development. This gene encodes a DNA methyltransferase which is thought to function in de novo methylation, rather than maintenance methylation. The protein localizes primarily to the nucleus and its expression is developmentally regulated. Mutations in this gene cause the immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome. Six alternatively spliced transcript variants have been described. The full length sequences of variants 4 and 5 have not been determined.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Epigenetic Regulation of CDH1 Is Altered after HOXB7-Silencing in MDA-MB-468 Triple-Negative Breast Cancer Cells.

Genes

Paço A,Leitão-Castro J,Freitas R

PA1-884 was used in Western Blot to explore the contribution of HOXB7 deregulation to the aggressiveness of certain cancers such as triple-negative breast cancers.

Sun Oct 03 00:00:00 EDT 2021

Epigenetic Regulation of CDH1 Is Altered after HOXB7-Silencing in MDA-MB-468 Triple-Negative Breast Cancer Cells.

Genes

Paço A,Leitão-Castro J,Freitas R

PA1-884 was used in Western Blot to explore the contribution of HOXB7 deregulation to the aggressiveness of certain cancers such as triple-negative breast cancers.

Sun Oct 03 00:00:00 EDT 2021

Decreases in different Dnmt3b activities drive distinct development of hematologic malignancies in mice.

The Journal of biological chemistry

Lopusna K,Nowialis P,Opavska J,Abraham A,Riva A,Haney SL,Opavsky R

Published figure using DNMT3B polyclonal antibody (Product # PA1-884) in Western Blot

Tue Aug 31 00:00:00 EDT 2021

Dnmt3b catalytic activity is critical for its tumour suppressor function in lymphomagenesis and is associated with c-Met oncogenic signalling.

EBioMedicine

Lopusna K,Nowialis P,Opavska J,Abraham A,Riva A,Opavsky R

PA1-884 was used in Western Blot to study a role of Dnmt3b's catalytic activity in development and cancer using a mouse model expressing catalytically inactive Dnmt3b (Dnmt3bCI).

Fri Jan 01 00:00:00 EST 2021

Dnmt3b catalytic activity is critical for its tumour suppressor function in lymphomagenesis and is associated with c-Met oncogenic signalling.

EBioMedicine

Lopusna K,Nowialis P,Opavska J,Abraham A,Riva A,Opavsky R

PA1-884 was used in Western Blot to study a role of Dnmt3b's catalytic activity in development and cancer using a mouse model expressing catalytically inactive Dnmt3b (Dnmt3bCI).

Fri Jan 01 00:00:00 EST 2021

Catalytically inactive Dnmt3b rescues mouse embryonic development by accessory and repressive functions.

Nature communications

Nowialis P,Lopusna K,Opavska J,Haney SL,Abraham A,Sheng P,Riva A,Natarajan A,Guryanova O,Simpson M,Hlady R,Xie M,Opavsky R

PA1-884 was used in Western Blotting to highlight that Dnmt3b is a multifaceted protein that serves as an enzyme, an accessory factor for other methyltransferases, and as a transcriptional repressor in mouse embryogenesis.

Thu Sep 26 00:00:00 EDT 2019

Diabetes impairs wound healing by Dnmt1-dependent dysregulation of hematopoietic stem cells differentiation towards macrophages.

Nature communications

Yan J,Tie G,Wang S,Tutto A,DeMarco N,Khair L,Fazzio TG,Messina LM

PA1-884 was used in Western Blotting to demonstrate for the first time that type 2 diabetes mellitus induces an haematopoietic stem cell-autonomous mechanism that causes impaired wound healing.

Tue Jan 02 00:00:00 EST 2018

The SNF2 family ATPase LSH promotes cell-autonomous de novo DNA methylation in somatic cells.

Nucleic acids research

Termanis A,Torrea N,Culley J,Kerr A,Ramsahoye B,Stancheva I

PA1-884 was used in western blot determine if the capacity for de novo DNA methylation of developmentally regulated loci is preserved in differentiated somatic cells

Mon Sep 19 00:00:00 EDT 2016

G9a/GLP Complex Maintains Imprinted DNA Methylation in Embryonic Stem Cells.

Cell reports

Zhang T,Termanis A,Özkan B,Bao XX,Culley J,de Lima Alves F,Rappsilber J,Ramsahoye B,Stancheva I

PA1-884 was used in western blot to characterize the G9a/GLP complex and how it maintains imprinted DNA methylation in embryonic stem cells

Tue Apr 05 00:00:00 EDT 2016

LSH and G9a/GLP complex are required for developmentally programmed DNA methylation.

Genome research

Myant K,Termanis A,Sundaram AY,Boe T,Li C,Merusi C,Burrage J,de Las Heras JI,Stancheva I

PA1-884 was used in western blot to investigate the role of LSH and G9a/GLP in DNA methylation during development

Sat Jan 01 00:00:00 EST 2011

De novo DNA methyltransferase DNMT3b interacts with NEDD8-modified proteins.

The Journal of biological chemistry

Shamay M,Greenway M,Liao G,Ambinder RF,Hayward SD

PA1-884 was used in western blot to study the interaction between de novo DNA methyltransferase DNMT3b and NEDD8-modified proteins

Fri Nov 19 00:00:00 EST 2010

LSH cooperates with DNA methyltransferases to repress transcription.

Molecular and cellular biology

Myant K,Stancheva I

PA1-884 was used in western blot to investigate the role of LSH during DNA methylation by DNA methyltransferases in mammals

Tue Jan 01 00:00:00 EST 2008

Bioinformatics

Protein Aliases: DNA (cytosine-5)-methyltransferase 3B; DNA (cytosine-5-)-methyltransferase 3 beta; DNA methyltransferase HsaIIIB; DNA methyltransferase MmuIIIB; DNA MTase HsaIIIB; DNA MTase MmuIIIB; Dnmt3; Dnmt3b; m.MmuIIIB; MGC1090

Gene Aliases: DNMT3B; ICF; ICF1; M.HsaIIIB; MmuIIIB

UniProt ID: (Human) Q9UBC3, (Mouse) O88509

Entrez Gene ID: (Human) 1789, (Mouse) 13436

Molecular Function: DNA methyltransferase