DNMT1 Antibody (60B1220.1)

Western Blot: DNMT1 Antibody (60B1220.1) [NB100-56519] - Analysis of Dnmt1 in 2102EP (human embryonic carcinoma) in the A) absence and B) presence of immunizing peptide, recombinant human Dnmt1 protein in the C) absence and D) presence of immunizing peptide, NIH 3T3 (mouse embryonic fribroblast) in the E) absence and F) presence of immunizing peptide, and D3 (mouse embryonic stem cell) in the G) absence and H) presence of immunizing peptide using Dnmt1 antibody. this antibody. 5 ug/ml for testing the human cell line and 0.1 ug/ml was used for testing the recombinant protein and the mouse cell lines. ’s goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used for this test.

Immunocytochemistry/Immunofluorescence: DNMT1 Antibody (60B1220.1) [NB100-56519] - Dnmt1 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red). Antibody dilution of 1:10 was used. Image objective 40x.

Immunohistochemistry-Paraffin: DNMT1 Antibody (60B1220.1) [NB100-56519] - Tissue section of human hepatocellular carcinoma at 1:50 dilution.

Flow Cytometry: DNMT1 Antibody (60B1220.1) [NB100-56519] - An intracellular stain was performed on HeLa cells with DNMT1 [60B1220.1] Antibody NB100-56519AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Flow Cytometry: DNMT1 Antibody (60B1220.1) [NB100-56519] - Analysis of PE conjugate of NB100-56519. An intracellular stain was performed on HeLa cells with DNMT1 antibody (60B1220.1) NB100-56519PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized with 0

Simple Western: DNMT1 Antibody (60B1220.1) [NB100-56519] - Simple Western lane view shows a specific band for Dnmt1 in 0.5 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Po, Bv, Sh
• Applications: WB, Simple Western, ChIP, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready, IHC-FrFl
• Clone: 60B1220.1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

DNMT1 Antibody (60B1220.1) Summary

• Immunogen: This antibody was raised against a synthetic peptide corresponding to amino acids 637-650 (EKDDREDKENAFKR) of human Dnmt1 (Genbank Accession No. NP_001370).
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: DNMT1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 - 0.5 ug/mL
  • Simple Western 1:100
  • Chromatin Immunoprecipitation 1:20 - 1:1000
  • Flow Cytometry 1 - 5 ug/mL
  • Immunocytochemistry/Immunofluorescence 1:10
  • Immunohistochemistry 1:50 - 1:500
  • Immunohistochemistry-Frozen 1:500
  • Immunohistochemistry-Paraffin 1:50 - 1:500
  • Immunoprecipitation 1:10 - 1:500
  • CyTOF-ready
  • Flow (Intracellular)
  • Immunohistochemistry Free-Floating 1:500
• Application Notes: Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 24386225). Use in Flow-intracellular reported in scientific literature (PMID: 24578214).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Control:

  DNMT1 Knockout 293T Cell Lysate

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 19924110). Porcine reactivity reported in scientific literature (PMID: 23808878). Sheep reactivity reported in scientific literature (PMID: 20413592). Bovine reactivity reported in scientific literature (PMID:27384909).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for DNMT1 Antibody (60B1220.1)

• AIM
• CXXC9
• CXXC9CXXC finger protein 9
• CXXC-type zinc finger protein 9
• DNA (cytosine-5-)-methyltransferase 1
• DNA (cytosine-5)-methyltransferase 1
• DNA methyltransferase 1
• DNA methyltransferase HsaI
• DNA MTase HsaI
• DNMT
• DNMT1
• EC 2.1.1.37
• FLJ16293
• m.HsaI
• MCMT
• MCMTMGC104992

Background

Methylation of DNA at cytosine residues plays an important role in regulation of gene expression, genomic imprinting and is essential for mammalian development. Hypermethylation of CpG islands in tumor suppressor genes or hypomethylation of bulk genomic DNA may be linked with development of cancer. To date, 3 families of mammalian DNA methyltransferase genes have been identified which include Dnmt1, Dnmt2 and Dnmt3. Dnmt1 is constitutively expressed in proliferating cells and inactivation of this gene causes global demethylation of genomic DNA and embryonic lethality. Dnmt2 is expressed at low levels in adult tissues and its inactivation does not affect DNA methylation or maintenance of methylation. The Dnmt3 family members, Dnmt3a and Dnmt3b, are strongly expressed in ES cells but their expression is down regulated in differentiating ES cells and is low in adult somatic tissue. Dnmt1 co-purifies with the retinoblastoma (Rb) tumour suppressor gene product, E2F1, and HDAC1. Dnmt1 also cooperates with Rb to repress transcription from promoters containing E2F-binding sites suggesting a link between DNA methylation, histone deacetylase and sequence-specific DNA binding activity, as well as a growth-regulatory pathway that is disrupted in nearly all cancer cells. NB100-56519 recognizes Dnmt1, a 1616 amino acid protein according to GenBank no. NP_001370.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for DNMT1 Antibody (NB100-56519)(114)

Publications using NB100-56519

• Sun X, Cui Y, Feng H et al. TGF-beta signaling controls Foxp3 methylation and T reg cell differentiation by modulating Uhrf1 activity J. Exp. Med. Sep 12 2019 [PMID: 31515281]
• Kuo HD, Wu R, Li S et al. Anthocyanin Delphinidin Prevents Neoplastic Transformation of Mouse Skin JB6 P+ Cells: Epigenetic Re-activation of Nrf2-ARE Pathway AAPS J Jun 28 2019 [PMID: 31254216]
• Fisslthaler B, Zippel N, Abdel Malik R et al. Myeloid-Specific Deletion of the AMPK alpha 2 Subunit Alters Monocyte Protein Expression and Atherogenesis Int J Mol Sci Jun 19 2019 [PMID: 31248224] (WB, Mouse)
• Gatta E, Grayson DR, Auta J et al. Genome-wide methylation in alcohol use disorder subjects: implications for an epigenetic regulation of the cortico-limbic glucocorticoid receptors (NR3C1) Mol. Psychiatry Jun 25 2019 [PMID: 31239533] (ChIP, Human)
• Huang M, Chou Y, Li M et al. Epigenetic Targeting DNMT1 of Pancreatic Ductal Adenocarcinoma using interstitial control release biodegrading polymer reduced tumor growth through Hedgehog pathway inhibition PLoS ONE Oct 16 2018 [PMID: 30385365] (IHC, Human)
• Miotto B, Marchal C, Adelmant G et al. Stabilization of the methyl-CpG binding protein ZBTB38 by the deubiquitinase USP9X limits the occurrence and toxicity of oxidative stress in human cells. Nucleic Acids Res. Feb 27 2018 [PMID: 29490077] (WB, Human)
• Meliso FM, Micali D, Silva CT et al. SIRT1 regulates Mxd1 during malignant melanoma progression Oncotarget 2017 Dec 29 [PMID: 29383100] (ChIP)
• Yang J, Wu R, Li W et al. The triterpenoid corosolic acid blocks transformation and epigenetically reactivates Nrf2 in TRAMP-C1 prostate cells. Mol. Carcinog. 2017 Dec 16 [PMID: 29247555] (Mouse)
• Li W, Su ZY, Guo Y et al. Curcumin Derivative Epigenetically Reactivates Nrf2 Antioxidative Stress Signaling in Mouse Prostate Cancer TRAMP C1 Cells. Chem. Res. Toxicol. 2017 Dec 11 [PMID: 29228771] (Mouse)
• Chu YY, Ko CY, Wang SM et al. Bortezomib-induced miRNAs direct epigenetic silencing of locus genes and trigger apoptosis in leukemia Cell Death Dis. 2017 Nov 09 [PMID: 29120412] (Human)
• Kuang H, Tang Z, Zhang C et al. Taxifolin Activates the Nrf2 Anti-Oxidative Stress Pathway in Mouse Skin Epidermal JB6 P+ Cells through Epigenetic Modifications Int J Mol Sci 2017 Jul 17 [PMID: 28714938] (WB, Human)
• Sheikh MA, Malik YS, Zhu X. RA-Induced Transcriptional Silencing of Checkpoint Kinase-2 through Promoter Methylation by Dnmt3b Is Required for Neuronal Differentiation of P19 Cells J. Mol. Biol. 2017 Jul 13 [PMID: 28712951] (Mouse)
• Miozzo F, Arnould H, de Thonel A et al. Alcohol exposure promotes DNA methyltransferase DNMT3A upregulation through reactive oxygen species-dependent mechanisms Cell Stress Chaperones 2017 Jul 15 [PMID: 28712054] (Mouse)
• He J, Pei L, Jiang H et al. Chemoresistance of colorectal cancer to 5-fluorouracil is associated with silencing of the BNIP3 gene through aberrant methylation. Journal of Cancer. Apr 9 2017 [PMID: 28607593] (WB, Human)
• Szarc Vel Szic K, Declerck K, Crans RAJ et al. Epigenetic silencing of triple negative breast cancer hallmarks by withaferin A. Oncotarget Apr 13 2017 [PMID: 28467815] (WB, Human)
• Anwar SL, Krech T, Hasemeier B et al. hsa-mir-183 is frequently methylated and related to poor survival in human hepatocellular carcinoma. World J. Gastroenterol. Mar 7 2017 [PMID: 28321157] (WB, Human)
• Raciti GA, Spinelli R, Desiderio A et al. Specific CpG hyper-methylation leads to Ankrd26 gene down-regulation in white adipose tissue of a mouse model of diet-induced obesity. Sci Rep. Mar 7 2017 [PMID: 28266632] (ChIP, Mouse)
• Neri F, Rapelli S, Krepelova A et al. Intragenic DNA methylation prevents spurious transcription initiation. Nature. Feb 22 2017 [PMID: 28225755] (WB, Human)
• Wilson RH, Biasutto AJ, Wang L et al. PCNA dependent cellular activities tolerate dramatic perturbations in PCNA client interactions DNA Repair (Amst.) Feb 1 2017 [PMID: 28073635] (WB, Human)
• Koh HB, Scruggs AM, Huang SK. Transforming Growth Factor-beta1 Increases DNA Methyltransferase 1 and -3a Expression Through Distinct Post-transcriptional Mechanisms in Lung Fibroblasts. J. Biol. Chem. Jul 12 2016 [PMID: 27405758] (IP, WB, ICC/IF, Human)
• Daniel FI, Alves SR, Vieira DS et al. Immunohistochemical expression of DNA methyltransferases 1, 3a, and 3b in actinic cheilitis and lip squamous cell carcinomas. J. Oral Pathol. Med. May 9 2016 [PMID: 27159259] (IHC-P, Human)
  
  Details:
  DNMT1 antibody (clone 60B1220.1; Imgenex IMG-261A) was used at 1:1500 dilution for IHC-P analysis of tissues from human subjects with actinic cheilitis and lip squamous cell carcinoma (LSCC), and non-neoplastic tissue (Figure 1).
• Li W Pung D, Su ZY Guo Y, Zhang C et al. Epigenetics Reactivation of Nrf2 in Prostate TRAMP C1 Cells by Curcumin Analogue FN1. ACS. 2016 Apr 18 [PMID: 26991801] (WB, Human)
• Hong B, Su Z, Zhang C et al. Reserpine Inhibit the JB6 P+ Cell Transformation Through Epigenetic Reactivation of Nrf2-Mediated Anti-oxidative Stress Pathway. AAPS J. 2016 Mar 17 [PMID: 26988984] (WB, Mouse)
• Miao CG, Qin D, Du CL et al. DNMT1 activates the canonical Wnt signaling in rheumatoid arthritis model rats via a crucial functional crosstalk between miR-152 and the DNMT1, MeCP2 Int. Immunopharmacol. 2015 Jun 17 [PMID: 26093272] (ChIP, Rat)
• Kopparapu PK, Miranda C, Fogelstrand L et al. MCPH1 Maintains long-term epigenetic silencing of angpt2 in chronic lymphocytic leukemia FEBS J. 2015 Feb 21 [PMID: 25703238] (Human)
• Neri F, Incarnato D, Krepelova A et al. Single-Base Resolution Analysis of 5-Formyl and 5-Carboxyl Cytosine Reveals Promoter DNA Methylation Dynamics Cell Rep. 2015 Feb 04 [PMID: 25660018] (WB, Mouse)
  
  Details:
  DNMT1 antibody (clone 60B1220.1) used for WB analysis of lysates from mouse embryonic stem cell / mESCs which were transfected with either control shRNA (shCtr) or shRNAs targeting Dnmts namely Sh-Dnmt1, Sh-Dnmt3a, Sh-Dnmt3b and Sh-Dnmt3a/b (Figure 4E).
• Guo Y, Shu L, Zhang C et al. Curcumin inhibits anchorage-independent growth of HT29 human colon cancer cells by targeting epigenetic restoration of the tumor suppressor gene DLEC1 Biochem. Pharmacol. 2015 Jan 29 [PMID: 25640947] (WB, Human)
  
  Details:
  DNMT1 antibody used for WB on lysates of HT29 human colon cancer cells treated or not with 2.5uM or 5uM Curcumin, and 2.5uM azadeoxycytidine/5AZA + 100uM trichostatin A/TSA. Curcumin was found to downregualte the expression levels of DNMT1 protein in a concentration-dependent manner (Fig 5).
• Huichalaf Claudia, Micheloni Stefano, Ferri Giulia et al. DNA methylation analysis of the macrosatellite repeat associated with FSHD muscular dystrophy at single nucleotide level. PLoS One. 2014 [PMID: 25545674] (ChIP)
• Bittencourt Danielle, Lee Brian H, Gao Lu et al. Role of distinct surfaces of the G9a ankyrin repeat domain in histone and DNA methylation during embryonic stem cell self-renewal and differentiation. Epigenetics Chromatin. 2014 [PMID: 25478012] (WB)
• Swiech L, Heidenreich M, Banerjee A et al. In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9. Nat. Biotechnol. 2014 Oct 19 [PMID: 25326897] (WB, Mouse)
• Wang L, Zhang C, Guo Y et al. Blocking of JB6 Cell Transformation by Tanshinone IIA: Epigenetic Reactivation of Nrf2 Antioxidative Stress Pathway. AAPS J. 2014 Nov 01 [PMID: 25274607] (WB, Mouse)
  
  Details:
  DNMT1 antibody used for WB on lysates of JB6 P+ cells (a mouse keratinocyte cell line) treated or not with Tanshinone IIA (TIIA), an active component in Salvia miltiorrhiza that has been reported to activate Nrf2 pathway.
• Fang M, Ou J, Hutchinson L, Green MR. The BRAF Oncoprotein Functions through the Transcriptional Repressor MAFG to Mediate the CpG Island Methylator Phenotype. Mol. Cell. 2014 Sep 18 [PMID: 25219500] (ChIP, IP, Human)
  
  Details:
  Dnmt1 antibody used for ChIP and CoIP applications in experiments involving DLD-1 (epithelial adherent cell line derived from Dukes' type C, colorectal adenocarcinoma), RKO colon carcinoma cells, SW48, VACO432 and PFFs cells.
• Guimaraes DM, Antunes DM, Duarte CM et al. DNA methyltransferase immunohistochemical expression in odontogenic tumours J. Oral Pathol. Med. 2014 Jul 24 [PMID: 25060239] (IHC-P, Human)
  
  Details:
  Dnmt1 antibody used for IHC-P at 1:400 dilution on formalin-fixed and paraffin-embedded tissue sections of human ameloblastomas, calcifying cystic odontogenic tumours, calcifying epithelial tumours, adenomatoid odontogenic tumours, keratocystic odontogenic tumours, ameloblastic fibromas, ameloblastic fibro-odontomas, central odontogenic fibromas, peripheral odontogenic fibromas and odontogenic myxomas.
• Fang P, Zhang D, Cheng Z et al. Hyperhomocysteinemia Potentiates Hyperglycemia-induced Inflammatory Monocyte Differentiation and Atherosclerosis Diabetes. 2014 Jul 09 [PMID: 25008174] (WB)
• Tajrishi MM, Shin J, Hetman M, Kumar A. DNA methyltransferase 3a and mitogen-activated protein kinase signaling regulate the expression of fibroblast growth factor-inducible 14 (Fn14) during denervation-induced skeletal muscle atrophy. J. Biol. Chem. 2014 Jun 03 [PMID: 24895120] (WB, ChIP, Mouse)
  
  Details:
  ChIP: Fig 5C (muscle); WB: Fig 5B (muscle).
• Anwar SL, Krech T, Hasemeier B et al. Deregulation of RB1 expression by loss of imprinting in human hepatocellular carcinoma. J. Pathol. 2014 May 16 [PMID: 24838394] (WB, Human)
  
  Details:
  HLE human hepatoma cells, Fig S7. The specificity of the DNMT1 mAb was validated with DNMT1-siRNA by WB, Fig S7. Note: The DNMT1 mAb was used to demonstrate that DNMT1 siRNA knocked down DNMT1 in the researcher's model system.
• Paredes-Gonzalez X, Fuentes F, Su ZY, Kong AN. Apigenin Reactivates Nrf2 Anti-oxidative Stress Signaling in Mouse Skin Epidermal JB6 P + Cells Through Epigenetics Modifications. AAPS J. 2014 May 16 [PMID: 24830944] (WB, Mouse)
  
  Details:
  JB6 P+ mouse skin epidermal cells, Fig 6a.
• Chuang C-H, Wang W-J, Li C-F et al. The combination of the prodrugs perforin-CEBPD and perforin-granzyme B efficiently enhances the activation of caspase signaling and kills prostate cancer. Cell Death Dis. 2014 [PMID: 24810056] (WB)
• Calabrese R, Valentini E, Ciccarone F et al. TET2 gene expression and 5-hydroxymethylcytosine level in multiple sclerosis peripheral blood cells. Biochim. Biophys. Acta 2014 Apr 13 [PMID: 24735979] (WB, Human)
  
  Details:
  PBMC, Fig 1D
• Serra FW, Fang M, Park SM et al. A KRAS-directed transcriptional silencing pathway that mediates the CpG island methylator phenotype. Elife 2014 Mar 13 [PMID: 24623306] (ChIP, Human)
• Neidhart M, Karouzakis E, Jungel A et al. Inhibition of spermidine/spermine N1-acetyltransferase (SSAT1) activity - a new therapeutical concept in rheumatoid arthritis. Arthritis & rheumatology. 2014 Feb 27 [PMID: 24578214] (Flow-IC, Human)
  
  Details:
  Primary human synovial fibroblasts isolated from patients with rheumatoid arthritis, Table 1, Fig 3B.
• Teneng I, Tellez CS, Picchi MA et al. Global identification of genes targeted by DNMT3b for epigenetic silencing in lung cancer. Oncogene. 2014 Jan 27 [PMID: 24469050] (ChIP, Human)
• Su ZY, Zhang C, Lee JH et al. Requirement and Epigenetics Reprogramming of Nrf2 in Suppression of Tumor Promoter TPA-Induced Mouse Skin Cell Transformation by Sulforaphane. Cancer Prev Res (Phila). 2014 Mar 1 [PMID: 24441674] (WB, Mouse)
  
  Details:
  JB6 P+ mouse epidermal cells, Fig 6.
• Hara ES1, Ono M1, Eguchi T2 et al. miRNA-720 Controls Stem Cell Phenotype, Proliferation and Differentiation of Human Dental Pulp Cells. PLoS One 2013 Dec 30 [PMID: 24386225] (ICC/IF, Human)
• Otani J, Kimura H, Sharif J et al. Cell Cycle-Dependent Turnover of 5-Hydroxymethyl Cytosine in Mouse Embryonic Stem Cells. PLoS One 2013 Dec 10 [PMID: 24340069] (ChIP, Mouse)
• Watson CJ, Collier P, Tea I et al. Hypoxia-induced epigenetic modifications are associated with cardiac tissue fibrosis and the development of a myofibroblast-like phenotype. Hum Mol Genet 2013 Dec 16 [PMID: 24301681] (WB, Human)
• Kanduri Meena, Sander Birgitta, Ntoufa Stavroula et al. A key role for EZH2 in epigenetic silencing of HOX genes in mantle cell lymphoma. Epigenetics. 2013 Dec [PMID: 24107828] (ChIP)
• Giraldo Angelica M, DeCourcy Kristi, Ball Suyapa F et al. Gene expression of Dnmt1 isoforms in porcine oocytes, embryos, and somatic cells. Cell Reprogram. 2013 Aug [PMID: 23808878] (ICC/IF, Porcine)
• Sheikh MA, Malik YS, Yu H et al. Epigenetic regulation of Dpp6 expression by Dnmt3b and its novel role in the inhibition of RA induced neuronal differentiation of P19 cells. PLoS One. 2013 [PMID: 23409053]
• Ip J, Canham P, Choo KH et al. Normal DNA methylation dynamics in DICER1-deficient mouse embryonic stem cells. PLoS Genet. 2012 Sep [PMID: 22969435]
• Kahlon N. The Effects of Oocyte Exposure to Bisphenol A On Early Bovine Embryo Development. Thesis. May 10 2016 (WB, ICC/IF, Bovine)
• Anwar SL, Albat C, Krech T et al. Concordant hypermethylation of intergenic microRNA genes in human hepatocellular carcinoma as new diagnostic and prognostic marker. Int J Cancer. 2013 Aug 1 [PMID: 23364900]
• Grau L, Luque-Garcia JL, Gonzalez-Peramato P et al. A quantitative proteomic analysis uncovers the relevance of CUL3 in bladder cancer aggressiveness. PLoS One. 2013 [PMID: 23308193]
• Anwar SL, Krech T, Hasemeier B et al. Loss of imprinting and allelic switching at the DLK1-MEG3 locus in human hepatocellular carcinoma. PLoS One. 2012 [PMID: 23145177]
• Ji W, Yang L, Yuan J et al. MicroRNA-152 targets DNA methyltransferase 1 in NiS-transformed cells via a feedback mechanism. Carcinogenesis. 2013 Feb [PMID: 23125218]
• Zimmermann N, Zschocke J, Perisic T et al. Antidepressants inhibit DNA methyltransferase 1 through reducing G9a levels. Biochem J. 2012 Nov 15 [PMID: 22880885]
• Mohammad F, Pandey GK, Mondal T et al. Long noncoding RNA-mediated maintenance of DNA methylation and transcriptional gene silencing. Development. 2012 Aug [PMID: 22721776]
• Bhardwaj A, Song HW, Beildeck M et al. DNA demethylation-dependent AR recruitment and GATA factors drive Rhox5 homeobox gene transcription in the epididymis. Mol Endocrinol. 2012 Apr [PMID: 22322598]
• Selvakumar T, Gjidoda A, Hovde SL, Henry RW. Regulation of human RNA polymerase III transcription by DNMT1 and DNMT3a DNA methyltransferases. J Biol Chem. 2012 Mar 2 [PMID: 22219193]
• Kadriu B, Guidotti A, Chen Y, Grayson DR. DNA methyltransferases1 (DNMT1) and 3a (DNMT3a) colocalize with GAD67-positive neurons in the GAD67-GFP mouse brain. J Comp Neurol. 2012 Jun 15 [PMID: 22134929] (WB, IHC-Fr, Mouse)
  
  Details:
  Antibodies cited: 1. DNMT1 (IMG-261A): WB (mouse brain cortical extracts), Table 1; IHC (frozen) (mouse brain), Figs 5, 6, 8B, 8C. 2. DNMT3A (IMG-268A): WB (mouse brain cortical extracts), Table 1; IHC (frozen) (mouse brain), Fig 7. Note: The specificitie
• Intrinsic and extrinsic regulation of DNA methylation during malignant transformation. Thesis. 2014 (ChIP)
• Bittencourt D, Lee BH, Gao L. Role of distinct surfaces of the G9a ankyrin repeat domain in histone and DNA methylation during embryonic stem cell self-renewal and differentiation. Epigenetics & Chromatin. 2014 (WB, Mouse)
  
  Details:
  DNMT1 antibody (clone 60B1220.1) used for WB on whole cell extracts from wild-type (TT2) mouse embryonic stem cells /mESCs, G9a-null mESCs or distinct clonal ESC lines (indicated by #1 and #2) stably expressing wild-type G9a (+WT) or G9a harboring the indicated point mutations in the ANK domain (Figure 1B).
• Yang J, Corsello TR, Ma Y. Stem cell gene SALL4 suppresses transcription through recruitment of DNA methyltransferases J Biol Chem 2012 Jan [PMID: 22128185] (WB, Human)
• Rochlin Ilia, Turbow David, Gomez Frank et al. Predictive mapping of human risk for West Nile virus (WNV) based on environmental and socioeconomic factors. PLoS One. 2011 [PMID: 21853103]
• Saito Shunsuke, Kato Jun, Hiraoka Sakiko et al. DNA methylation of colon mucosa in ulcerative colitis patients: correlation with inflammatory status. Inflamm Bowel Dis. 2011 Sep [PMID: 21830274] (Human)
• Fan Huitao, Cinar Mehmet Ulas, Phatsara Chirawath et al. Molecular mechanism underlying the differential MYF6 expression in postnatal skeletal muscle of Duroc and Pietrain breeds. Gene. 2011 Oct 15 [PMID: 21749918] (WB)
• Dhawan Sangeeta, Georgia Senta, Tschen Shuen-Ing et al. Pancreatic beta cell identity is maintained by DNA methylation-mediated repression of Arx. Dev Cell. 2011 Apr 19 [PMID: 21497756] (ICC/IF, Mouse)
• Nasonkin Igor O, Lazo Kevin, Hambright Dustin et al. Distinct nuclear localization patterns of DNA methyltransferases in developing and mature mammalian retina. J Comp Neurol. 2011 Jul 1 [PMID: 21452232] (WB, Mouse)
• Molognoni Fernanda, Cruz Adriana T, Meliso Fabiana M et al. Epigenetic reprogramming as a key contributor to melanocyte malignant transformation. Epigenetics. 2011 Apr [PMID: 21343701] (WB, Mouse)
• Yamanaka Ken-ichi, Sakatani Miki, Kubota Kaiyu et al. Effects of downregulating DNA methyltransferase 1 transcript by RNA interference on DNA methylation status of the satellite I region and in vitro development of bovine somatic cell nuclear transfer embryos. J Reprod Dev. 2011 Jun [PMID: 21343670]
• Dalvai M, Mondesert O, Bourdon J-C et al. Cdc25B is negatively regulated by p53 through Sp1 and NF-Y transcription factors. Oncogene. 2011 May 12 [PMID: 21242964] (Human)
• Teneng Ivo, Montoya-Durango Diego E, Quertermous James L et al. Reactivation of L1 retrotransposon by benzo(a)pyrene involves complex genetic and epigenetic regulation. Epigenetics. 2011 Mar [PMID: 21150308] (WB)
• Daniel Filipe Ivan, Rivero Elena Riet Correa, Modolo Filipe et al. Immunohistochemical expression of DNA methyltransferases 1, 3a and 3b in oral leukoplakias and squamous cell carcinomas. Arch Oral Biol. 2010 Dec [PMID: 20951977] (Human)
• Pierard V, Guiguen A, Colin L et al. DNA cytosine methylation in the bovine leukemia virus promoter is associated with latency in a lymphoma-derived B-cell line: potential involvement of direct inhibition of cAMP-responsive element (CRE)-binding protein/CRE modulator/activation transcription J Biol Chem. 2010 Jun 18 [PMID: 20413592] (WB, Sheep)
  
  Details:
  western blot: DNMT1 (IMG-261A), DMNT3A (IMG-268A), DNMT3B (IMG-184A) in Fig 8: Nuclear extracts from ovine (sheep) cell lines L267 and L267LTaxSN. L267 is a B-cell lymphoma cell line established from Bovine Leukemia Virus (BLV) infected sheep. L267LTaxSN is a derivative of L267, see the publication for additional details.
• Wu Y, Alvarez M, Slamon DJ et al. Caspase 8 and maspin are downregulated in breast cancer cells due to CpG site promoter methylation. BMC Cancer. 2010 Feb 4 [PMID: 20132554] (WB, Human)
  
  Details:
  Dnmt1 (IMG-261A),Dnmt2 (IMG-281), Dnmt3a (IMG-268A), Dnmt3b (IMG-184A). WB: Human breast cancer MCF-7, MB231, BT474 cell lines treated with or without 5-aza-dc, Fig 4.
• Perisic Tatjana, Zimmermann Nicole, Kirmeier Thomas et al. Valproate and amitriptyline exert common and divergent influences on global and gene promoter-specific chromatin modifications in rat primary astrocytes. Neuropsychopharmacology. 2010 Feb [PMID: 19924110] (WB, Rat)
• Ye Ying, McDevitt Michael A, Guo Mingzhou et al. Progressive chromatin repression and promoter methylation of CTNNA1 associated with advanced myeloid malignancies. Cancer Res. 2009 Nov 1 [PMID: 19826047] (ChIP, Human)
• Hwang Cheol Kyu, Song Kyu Young, Kim Chun Sung et al. Epigenetic programming of mu-opioid receptor gene in mouse brain is regulated by MeCP2 and Brg1 chromatin remodelling factor. J Cell Mol Med. 2009 Sep [PMID: 19602036] (IP, Mouse)
• Zampieri M, Passananti C, Calabrese R et al. Parp1 localizes within the Dnmt1 promoter and protects its unmethylated state by its enzymatic activity. PLoS One. 2009 [PMID: 19262751] (WB, Mouse)
  
  Details:
  L929 mouse fibroblasts: WB (Figs 2A, 5B), IP (Fig 4D).
• Yamagata Y, Asada H, Tamura I et al. DNA methyltransferase expression in the human endometrium: down-regulation by progesterone and estrogen. Hum Reprod. 2009 May [PMID: 19202141] (IHC-P)
  
  Details:
  1. IMG-261A (dnmt1): IHC(parafin), human endometrium tissue, Fig. 1A. 2. IMG-184A (dnmt3b): IHC(parafin), human endometrium tissue, Fig. 1B.
• Lin HJ, Zuo T, Lin CH et al. Breast cancer-associated fibroblasts confer AKT1-mediated epigenetic silencing of Cystatin M in epithelial cells. Cancer Res. 2008 Dec 15 [PMID: 19074894] (ChIP, Human)
  
  Details:
  ChIP (human breast cancer cell line MCF10A), Fig. 5A.
• Ko CY, Hsu HC, Shen MR et al. Epigenetic silencing of CCAAT/enhancer-binding protein delta activity by YY1/polycomb group/DNA methyltransferase complex. J Biol Chem. 2008 Nov 7 [PMID: 18753137] (WB)
  
  Details:
  HeLa cells in Fig. 7A (IF), 7B (WB following a DNA Affinity Precipitation Assay of nuclear extracts), 7C and D (ChIP): 1. IMG-261A (Dnmt1) 2. IMG-268A (Dnmt3a) 3. IMG-184A (Dnmt3b).
• O'Hagan Heather M, Mohammad Helai P, Baylin Stephen B. Double strand breaks can initiate gene silencing and SIRT1-dependent onset of DNA methylation in an exogenous promoter CpG island. PLoS Genet. 2008 [PMID: 18704159] (IP, Human)
• Guastafierro T, Cecchinelli B, Zampieri M et al. CCCTC-binding factor activates PARP-1 affecting DNA methylation machinery. J Biol Chem. 2008 Aug 8 [PMID: 18539602] (WB, Mouse)
  
  Details:
  WB: (mouse fibroblasts), Figs. 2A, 4A; IP: (mouse fibroblasts), Fig. 2E.
• Xia G, Schneider-Stock R, Diestel A et al. Helicobacter pylori regulates p21(WAF1) by histone H4 acetylation. Biochem Biophys Res Commun. 2008 May 2 [PMID: 18302936] (WB, Human)
  
  Details:
  Hhuman NCI N87 gastric carcinoma cells: WB (Fig 1) 1. Dnmt1 (IMG-261) 2. Dnmt3a (IMG-268) 3. Dnmt3b (IMG-184)
• Russell D Fischer, Betts D H. Alternative splicing and expression analysis of bovine DNA methyltransferase 1. Dev Dyn. 2008 Apr [PMID: 18297739]
• Hodge David R, Cho Edward, Copeland Terry D et al. IL-6 enhances the nuclear translocation of DNA cytosine-5-methyltransferase 1 (DNMT1) via phosphorylation of the nuclear localization sequence by the AKT kinase. Cancer Genomics Proteomics. 2007 Nov-Dec [PMID: 18204201] (IHC, Human)
• Ji W, Yang L, Yu L et al. Epigenetic silencing of O6-methylguanine DNA methyltransferase gene in NiS-transformed cells. Carcinogenesis. 2008 Jun [PMID: 18204074] (ChIP, Human)
  
  Details:
  ChIP (human bronchial epethial cell line 16HBE cells), Fig. 6 A,B.
• Gazin Claude, Wajapeyee Narendra, Gobeil Stephane et al. An elaborate pathway required for Ras-mediated epigenetic silencing. Nature. 2007 Oct 25 [PMID: 17960246] (IP, Mouse)
• Achour M, Jacq X, Ronde P et al. The interaction of the SRA domain of ICBP90 with a novel domain of DNMT1 is involved in the regulation of VEGF gene expression. Oncogene. 2008 Apr 3 [PMID: 17934516] (WB)
  
  Details:
  WB: Fig 3a (Jurkat cells), 4a (human vascular smooth muscle HTVs-SM1 cells), 7A (Jurkat cells treated with DNMT1 or scrambled siRNA). IP/WB: Fig 3b (Jurkat cells), 4b (HTVs-SM1 cells). Note: DNMT1 was detected at ~180 kDa. Proliferating Jurkat cell cultur
• Mabaera R, Greene MR, Richardson CA et al. Neither DNA hypomethylation nor changes in the kinetics of erythroid differentiation explain 5-azacytidine's ability to induce human fetal hemoglobin. Blood. 2008 Jan 1 [PMID: 17916742] (WB, Human)
  
  Details:
  WB (human primary erythroid cells): Figs. 5C, 6B; The antibody is DNMT1 siRNA validated in both figures.
• Villa R, Pasini D, Gutierrez A et al. Role of the polycomb repressive complex 2 in acute promyelocytic leukemia. Cancer Cell. 2007 Jun [PMID: 17560333]
• Oh Bong-Kyeong, Kim Haeryoung, Park Hye-Jung et al. DNA methyltransferase expression and DNA methylation in human hepatocellular carcinoma and their clinicopathological correlation. Int J Mol Med. 2007 Jul [PMID: 17549390] (Human)
• Park IY, Sohn BH, Yu E et al. Aberrant epigenetic modifications in hepatocarcinogenesis induced by hepatitis B virus X protein. Gastroenterology. 2007 Apr [PMID: 17408664] (WB)
  
  Details:
  (Chang liver and Hep G2 cell lines): 1. IMG-261(DNMT1): WB, Figs. 1A, 4A,B. 2. IMG-268A (DNMT3A): WB, ChIP, Figs. 1B, 2B, 4A,B.
• Humar B, Fukuzawa R, Blair V et al. Destabilized adhesion in the gastric proliferative zone and c-Src kinase activation mark the development of early diffuse gastric cancer. Cancer Res. 2007 Mar 15 [PMID: 17363565]
• MacDonald JL, Gin CS, Roskams AJ. Stage-specific induction of DNA methyltransferases in olfactory receptor neuron development. Dev Biol. 2005 Dec 15 [PMID: 16288735] (IHC-P)
• Lin TS, Lee H, Chen RA et al. An association of DNMT3b protein expression with P16INK4a promoter hypermethylation in non-smoking female lung cancer with human papillomavirus infection. Cancer Lett. 2005 Aug 8 [PMID: 16004934] (IHC-P)
  
  Details:
  Dnmt1 (IMG-261A) [IHC-P, Fig.1 (lung tumors)].
• Srivastava BI, Srivastava MD. Establishment and characterization of SRIK-NKL: a novel CD8+ natural killer/T cell line derived from a patient with leukemic phase of acute lymphoblastic lymphoma. Leuk Res. 2005 Jul [PMID: 15927673]
• Ghoshal K, Datta J, Majumder S et al. 5-Aza-deoxycytidine induces selective degradation of DNA methyltransferase 1 by a proteasomal pathway that requires the KEN box, bromo-adjacent homology domain, and nuclear localization signal. Mol Cell Biol. 2005 Jun [PMID: 15899874] (ICC/IF)
• Park Hye-Jung, Yu Eunsil, Shim Yhong-Hee. DNA methyltransferase expression and DNA hypermethylation in human hepatocellular carcinoma. Cancer Lett. 2006 Feb 28 [PMID: 15885882] (Human)
• Srivastava MD, Alexander F, Tuthill RJ. Immunology of cutaneous vasculitis associated with both etanercept and infliximab. Scand J Immunol. 2005 Apr [PMID: 15853915]
• Leu YW, Yan PS, Fan M et al. Loss of estrogen receptor signaling triggers epigenetic silencing of downstream targets in breast cancer. Cancer Res. 2004 Nov. [PMID: 15548683] (ChIP)
• Park IY, Sohn BH, Choo JH et al. Deregulation of DNA methyltransferases and loss of parental methylation at the insulin-like growth factor II (Igf2)/H19 loci in p53 knockout mice prior to tumor development. J Cell Biochem. 2005 Feb 15 [PMID: 15543560] (WB, Mouse)
  
  Details:
  WB (Fig 1), mouse thymus and liver: DMNT1 (IMG-261A) DNMT3a (IMG-268A).
• Suzuki M, Sunaga N, Shames DS et al. RNA interference-mediated knockdown of DNA methyltransferase 1 leads to promoter demethylation and gene re-expression in human lung and breast cancer cells. Cancer Res. 2004 May 1 [PMID: 15126351] (WB, Human)
  
  Details:
  WB (Fig 1): NCI-H1299 non small cell lung cancer and HCC1954 breast cancer human cell lines. Cells treated with DNMT1 siRNA showed a dramatic reduction of DNMT1 expression. Note: The specificity of the DNMT1 (IMG-261A) was siRNA validated in Fig 1 by WB i
• Xin H, Yoon HG, Singh PB et al. Components of a pathway maintaining histone modification and heterochromatin protein 1 binding at the pericentric heterochromatin in Mammalian cells. J Biol Chem. 2004 Mar 5 [PMID: 14665632]
• Leu YW, Rahmatpanah F, Shi H et al. Double RNA interference of DNMT3b and DNMT1 enhances DNA demethylation and gene reactivation. Cancer Res. 2003 Oct 1 [PMID: 14559786]
• Craig JM, Earle E, Canham P et al. Analysis of mammalian proteins involved in chromatin modification reveals new metaphase centromeric proteins and distinct chromosomal distribution patterns. Hum Mol Genet. 2003 Dec 1 [PMID: 14519686]
• Choi MS, Shim YH, Hwa JY et al. Expression of DNA methyltransferases in multistep hepatocarcinogenesis. Hum Pathol. 2003 Jan [PMID: 12605361] (ICC/IF)
  
  Details:
  IF/ICC: HepG2 cell stained with Dnmt1 (IMG-261A) and Dnmt3a (IMG-268A), Fig 1.
• Kimura F, Seifert HH, Florl AR et al. Decrease of DNA methyltransferase 1 expression relative to cell proliferation in transitional cell carcinoma. Int J Cancer. 2003 May 1 [PMID: 12594811] (WB, Human)
  
  Details:
  WB: human bladder transitional cell carcinoma cell lines and primary cultured normal human uropithelial cells (Fig. 2).
• Alcock RA, Dey S, Chendil D et al. Farnesyltransferase inhibitor (L-744,832) restores TGF-beta type II receptor expression and enhances radiation sensitivity in K-ras mutant pancreatic cancer cell line MIA PaCa-2. Oncogene. 2002 Nov 7 [PMID: 12420225]
• Santoro R, Li J, Grummt I. The nucleolar remodeling complex NoRC mediates heterochromatin formation and silencing of ribosomal gene transcription. Nat Genet. 2002 Nov [PMID: 12368916]
• Tsai CN, Tsai CL, Tse KP et al. The Epstein-Barr virus oncogene product, latent membrane protein 1, induces the downregulation of E-cadherin gene expression via activation of DNA methyltransferases. Proc Natl Acad Sci U S A. 2002 Jul 23 [PMID: 12110730]
• Zhang Z, Deng C, Lu Q, Richardson B. Age-dependent DNA methylation changes in the ITGAL (CD11a) promoter. Mech Ageing Dev. 2002 May [PMID: 12020947]
• Dennis K, Fan T, Geiman T et al. Lsh, a member of the SNF2 family, is required for genome-wide methylation. Genes Dev. 2001 Nov 15 [PMID: 11711429]

Reviews for DNMT1 Antibody (NB100-56519) (4)

Average Rating: 4.5

(Based on 4 reviews)

We have 4 reviews tested in 1 species: Human.

reviewed by: Anonymous WB Human 01/30/2019

Summary

• Application: Western Blot
• Sample Tested: cell
• Species: Human
• Lot: A-3

reviewed by: Yichi Xu ChIP Human 01/30/2019

Summary

• Application: Chromatin Immunoprecipitation
• Sample Tested: cell
• Species: Human
• Lot: A-3

Comments

• Comments: More than 1×10e6 cells and 2.4 μg antibody are optimal.

reviewed by: Anonymous WB Human 04/14/2016

Summary

• Application: Western Blot
• Sample Tested: Prostate cancer cell lines, LnCAP
• Species: Human

Blocking

• Blocking Details: 5% BSA, 25 Celsius, 1 hr

Details

• Detection Notes: chemiluminescence, bio rad chem doc, exposure time 30seconds, actin control

Comments

• Comments: Worked great with no problems.

reviewed by: Anonymous IP Human 12/12/2014

Summary

• Application: Immunoprecipitation
• Sample Tested: See PMID 24163373
• Species: Human

Blocking

• Blocking Details: See PMID 24163373

Details

• Detection Notes: See PMID 24163373

Comments

• Comments: Published in PMID: 24163373

FAQs for DNMT1 Antibody (NB100-56519). (Showing 1 - 2 of 2 FAQs).

1. Could you please tell me the size of your DNMT1 protein along with Expression System? I am also doing mutant DNMT1 protein expression using the truncated part (650-1602=952 aa), using bacterial expression system (E.coli, BL21(De3). But right now I am getting some problem that, my size of protein is little higher than my accurate size? I use Auto-induction method to express the protein.
   • As you can see from our testing data, we see bands at around 185-200 kDa. Full details of the samples can be found on the image captions. Dnmt1,UniProt P26358, is also highly phosphorylated and acetylated which can raise the apparent molecular weight to higher than the predicted molecular weight. This is why you are seeing the change in size compared to what you might expect from your truncated protein.
2. We have recently purchased the above mentioned anti-DNMT1 antibody. However, the product datasheet does not have any exact details on the molecular weight of the antibody and images provided are with very wide range protein marker (near DNMT-1 band 116-200kD).
   • According to Uniprot #P26358 the theoretical molecular weight of human DNMT1 is 183 kDa for isoform 1, ~185 kDa for isoform 2 and 144 kDa for isoform 3. This antibody is designed to detect all 3 of these isoforms.Please not that this protein is subject to multiple posttranslational modifications which may differ by the species, cell type and disease state and these may affect how the protein runs on WB. In case of the WB image (image 1 of 5) we show several human an mouse cell lines, human recombinant protein, and each blot shows at least 1 band which looks like the predicted molecular weight of the target. I agree that the 116-200 range is rather wide and it is impossible to say that the protein runs exactly at 183kDa, but we are confident this is a specific band as, in addition, it is blocked by the blocking peptide.  The entire WB image is displayed to show that there were other protein bands observed in some cell lines, also blocked by the peptide. These may be similar epitopes present at other targets or results of posttranslational processing of the DNMT1 protein in these cell types. Further study would be necessary to determine what exactly are these, but we present this data to make the customers aware that there may be some additional bands on the WB.  If you are running a WB and ar enot confident that what you see is your target, we recommend to include some positive (overexpression lysate or treatment that induces the protein expression) or/and negative (KO, siRNA, cell line or tissue known not to express your target protein) controls.

Bioinformatics

• Gene Symbol: DNMT1
• Entrez:
  • 1786(Human)
  • 13433(Mouse)
• Uniprot:
  • P26358(Human)