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Novus Biologicals mouse monoclonal antibodies against dnmt3a
Fig. 2. Immunological detection of <t>DNMT3A</t> protein in seminoma (SE), non-seminoma (NS) and adja

Fig. 2. Immunological detection of <t>DNMT3A</t> protein in seminoma (SE), non-seminoma (NS) and adja

Mouse Monoclonal Antibodies Against Dnmt3a, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 179 article reviews

mouse monoclonal antibodies against dnmt3a - by Bioz Stars, 2026-03

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1) Product Images from "Up-regulation of DNA-methyltransferase 3A expression is associated with hypomethylation of intron 25 in human testicular germ cell tumors."

Article Title: Up-regulation of DNA-methyltransferase 3A expression is associated with hypomethylation of intron 25 in human testicular germ cell tumors.

Journal: The Tohoku journal of experimental medicine

doi: 10.1620/tjem.212.177

Figure Legend Snippet: Fig. 2. Immunological detection of DNMT3A protein in seminoma (SE), non-seminoma (NS) and adja

Techniques Used:

Figure Legend Snippet: Fig. 3. Analyses of DNMT3A methylation status. A: Methylation status of CpGs in the sequences

Techniques Used: Methylation

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Journal: The Tohoku journal of experimental medicine

Article Title: Up-regulation of DNA-methyltransferase 3A expression is associated with hypomethylation of intron 25 in human testicular germ cell tumors.

doi: 10.1620/tjem.212.177

Figure Lengend Snippet: Fig. 2. Immunological detection of DNMT3A protein in seminoma (SE), non-seminoma (NS) and adja

Article Snippet: The primary antibodies used were 200- and 400-fold dilutions of mouse monoclonal antibodies against DNMT3A (64B1446, Imgenex Co., San Diego, CA, USA) and γ - tubulin (GTU-88, Sigma-Aldrich, St. Louis, MO, USA), respectively.

Techniques:

Journal: The Tohoku journal of experimental medicine

Article Title: Up-regulation of DNA-methyltransferase 3A expression is associated with hypomethylation of intron 25 in human testicular germ cell tumors.

doi: 10.1620/tjem.212.177

Figure Lengend Snippet: Fig. 3. Analyses of DNMT3A methylation status. A: Methylation status of CpGs in the sequences

Techniques: Methylation

Journal: Oncology Reports

Article Title: Crosstalk between histone modification and DNA methylation orchestrates the epigenetic regulation of the costimulatory factors, Tim-3 and galectin-9, in cervical cancer

doi: 10.3892/or.2019.7388

Figure Lengend Snippet: Primer sequences.

Article Snippet: The antibodies used were as follows: Rabbit polyclonal antibody against human Tim-3 (1:500 dilution, ab185703; Abcam), rabbit polyclonal antibody against human galectin-9 (1:100 dilution, ab123712; Abcam), mouse monoclonal antibody against human DNMT3A (1:200 dilution, sc-365769; Santa Cruz Biotechnology), rabbit monoclonal antibody against human EZH2 (1:1,000 dilution, 5246; Cell Signaling Technology), rabbit monoclonal antibody against human H3K27me3 (1:1,000 dilution, 9733; Cell Signaling Technology), mouse monoclonal antibody against HPV18 E6 (1:250 dilution, NB100-2729; Novus Biologicals) and mouse monoclonal antibody against HPV18 E7 (1:250 dilution, NB110-17215; Novus Biologicals), β-actin mouse monoclonal antibody (1:500 dilution, 60008-1-lg; Proteintech).

Techniques: Sequencing, Control

DNMT3A regulates the expression of Tim-3/galectin-9 in cervical cancer cell lines. (A and B) DNMT3A-specific siRNA was transfected into HeLa cells, and the expression of DNMT3A and Tim-3/galectin-9 was detected by western blot analysis. β-actin protein was used as a loading control between lanes. (C) MS-PCR analysis was used to examine the methylation level of HAVCR2/LGALS9 in HeLa cells in which DNMT3A was knocked down. (D) Gray level analysis of HAVCR2/LGALS9 methylation levels in HeLa cells in which DNMT3A was knocked down (M, methylated; U, unmethylated); the methylated and unmethylated levels were quantified as M/M+U% and U/M+U%, respectively. (E and G) Schematic representation of the 4 regions of the HAVCR2/LGALS9 promoters amplified in the chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) experiment. (F and H) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-DNMT3A ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for the indicated 4 regions in the HAVCR2/LGALS9 promoters in qPCR. The enrichment of DNMT3A on HAVCR2/LGALS9 promoters relative to IgG in HeLa cells, and H3 against RPL30 was used as a positive control. *P<0.05; **P<0.01.

Journal: Oncology Reports

Article Title: Crosstalk between histone modification and DNA methylation orchestrates the epigenetic regulation of the costimulatory factors, Tim-3 and galectin-9, in cervical cancer

doi: 10.3892/or.2019.7388

Figure Lengend Snippet: DNMT3A regulates the expression of Tim-3/galectin-9 in cervical cancer cell lines. (A and B) DNMT3A-specific siRNA was transfected into HeLa cells, and the expression of DNMT3A and Tim-3/galectin-9 was detected by western blot analysis. β-actin protein was used as a loading control between lanes. (C) MS-PCR analysis was used to examine the methylation level of HAVCR2/LGALS9 in HeLa cells in which DNMT3A was knocked down. (D) Gray level analysis of HAVCR2/LGALS9 methylation levels in HeLa cells in which DNMT3A was knocked down (M, methylated; U, unmethylated); the methylated and unmethylated levels were quantified as M/M+U% and U/M+U%, respectively. (E and G) Schematic representation of the 4 regions of the HAVCR2/LGALS9 promoters amplified in the chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) experiment. (F and H) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-DNMT3A ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for the indicated 4 regions in the HAVCR2/LGALS9 promoters in qPCR. The enrichment of DNMT3A on HAVCR2/LGALS9 promoters relative to IgG in HeLa cells, and H3 against RPL30 was used as a positive control. *P<0.05; **P<0.01.

Techniques: Expressing, Transfection, Western Blot, Control, Methylation, Amplification, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Positive Control

EZH2 and H3K27me3 regulate Tim-3/galectin-9 expression through DNMT3A. (A) The protein levels of EZH2, H3K27me3 and DNMT3A in cervical cancer (Ca), paracarcinoma (Cap) (n=24) and normal cervical tissues (NC) (n=16) detected by western blot analysis. Blot images of 4 representative samples are shown from each group. (B) Densitometric analysis of EZH2, H3K27me3 and DNMT3A protein levels in cervical cancer (n=24), paracarcinoma (n=24) and normal cervical tissues (n=16). (C and F) Western blot analysis of HeLa cells 48 h following transfection with siRNA against EZH2. β-actin protein was used as a loading control between lanes. (D) MS-PCR analysis was used to examine the methylation level of HAVCR2/LGALS9 . (E) Gray level analysis of HAVCR2/LGALS9 methylation levels in HeLa cells in which EZH2 was knocked down (M, methylated; U, unmethylated); the methylated and unmethylated levels were quantified as M/M+U% and U/M+U%, respectively. (G) Schematic representation of the 4 regions of the DNMT3A promoter amplified in the chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) experiment. (H and I) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-EZH2 and anti-H3K27me3 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the DNMT3A promoters in qPCR. The enrichment of EZH2 and H3K27me3 on DNMT3A promoter relative to IgG in HeLa cells, and H3 against RPL30 was used as a positive control. **P<0.01; ns, not significant.

Journal: Oncology Reports

Article Title: Crosstalk between histone modification and DNA methylation orchestrates the epigenetic regulation of the costimulatory factors, Tim-3 and galectin-9, in cervical cancer

doi: 10.3892/or.2019.7388

Figure Lengend Snippet: EZH2 and H3K27me3 regulate Tim-3/galectin-9 expression through DNMT3A. (A) The protein levels of EZH2, H3K27me3 and DNMT3A in cervical cancer (Ca), paracarcinoma (Cap) (n=24) and normal cervical tissues (NC) (n=16) detected by western blot analysis. Blot images of 4 representative samples are shown from each group. (B) Densitometric analysis of EZH2, H3K27me3 and DNMT3A protein levels in cervical cancer (n=24), paracarcinoma (n=24) and normal cervical tissues (n=16). (C and F) Western blot analysis of HeLa cells 48 h following transfection with siRNA against EZH2. β-actin protein was used as a loading control between lanes. (D) MS-PCR analysis was used to examine the methylation level of HAVCR2/LGALS9 . (E) Gray level analysis of HAVCR2/LGALS9 methylation levels in HeLa cells in which EZH2 was knocked down (M, methylated; U, unmethylated); the methylated and unmethylated levels were quantified as M/M+U% and U/M+U%, respectively. (G) Schematic representation of the 4 regions of the DNMT3A promoter amplified in the chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) experiment. (H and I) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-EZH2 and anti-H3K27me3 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the DNMT3A promoters in qPCR. The enrichment of EZH2 and H3K27me3 on DNMT3A promoter relative to IgG in HeLa cells, and H3 against RPL30 was used as a positive control. **P<0.01; ns, not significant.

Techniques: Expressing, Western Blot, Transfection, Control, Methylation, Amplification, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Positive Control

Function of HPV18 oncoproteins E6/E7 in the regulation of Tim-3/galectin-9 and DNMT3A expression in human cervical cancer cells. (A) and (B) mRNA levels in of HPV18 E6/E7 -overexpressing C33A cells and of HPV18 E6/E7 -knockdown HeLa cells. (C) and (D) C33A cells were transfected with 18E6/E7/E6/7 lentiviral vector, and HeLa cells were transfected with 18E6/E7/E6/7-specific siRNAs. Western blot analysis was then used to detect DNMT3A expression. (E) Co-IP assay was used to analyzed the incorporation between DNMT3A and HPV18 E6/E7. (F and G) The methylation level of the HAVCR2/LGALS9 promoters was monitored by MS-PCR. Gray level analysis of HAVCR2/LGALS9 methylation levels in cervical cancer cells (M, methylated; U, unmethylated); the methylated and unmethylated levels were quantified as M/M+U% and U/M+U%, respectively. (H and I) C33A cells were transfected with 18E6/E7/E6/7 lentiviral vector, and HeLa cells were transfected with 18E6/E7/E6/7-specific siRNAs. The expression of Tim-3/galectin-9 was determined by western blot analysis. **P<0.01.

Journal: Oncology Reports

Article Title: Crosstalk between histone modification and DNA methylation orchestrates the epigenetic regulation of the costimulatory factors, Tim-3 and galectin-9, in cervical cancer

doi: 10.3892/or.2019.7388

Figure Lengend Snippet: Function of HPV18 oncoproteins E6/E7 in the regulation of Tim-3/galectin-9 and DNMT3A expression in human cervical cancer cells. (A) and (B) mRNA levels in of HPV18 E6/E7 -overexpressing C33A cells and of HPV18 E6/E7 -knockdown HeLa cells. (C) and (D) C33A cells were transfected with 18E6/E7/E6/7 lentiviral vector, and HeLa cells were transfected with 18E6/E7/E6/7-specific siRNAs. Western blot analysis was then used to detect DNMT3A expression. (E) Co-IP assay was used to analyzed the incorporation between DNMT3A and HPV18 E6/E7. (F and G) The methylation level of the HAVCR2/LGALS9 promoters was monitored by MS-PCR. Gray level analysis of HAVCR2/LGALS9 methylation levels in cervical cancer cells (M, methylated; U, unmethylated); the methylated and unmethylated levels were quantified as M/M+U% and U/M+U%, respectively. (H and I) C33A cells were transfected with 18E6/E7/E6/7 lentiviral vector, and HeLa cells were transfected with 18E6/E7/E6/7-specific siRNAs. The expression of Tim-3/galectin-9 was determined by western blot analysis. **P<0.01.

Techniques: Expressing, Knockdown, Transfection, Plasmid Preparation, Western Blot, Co-Immunoprecipitation Assay, Methylation

HPV18 oncoprotein E6/E7 alters the expression EZH2 and H3K27me3 in cervical cancer cells. (A and B) C33A cells were transfected with 18E6/E7/E6/7 lentiviral vector and HeLa cells were transfected with 18E6/E7/E6/7 specific siRNAs and western blot analysis was then used to detect EZH2 and H3K27me3 expression. (C) EZH2 and H3K27me3 expression in HeLa and C33A cells detected by western blot analysis. (D) Western blot analysis of the HPV18 E6/E7 protein level in HeLa cells following transfection with siRNAs against EZH2. (E) Co-IP assay was used to analyzed the incorporation between EZH2 and HPV18 E6/E7. (F) Schematic representation of the 4 regions of the EZH2 promoter amplified in the chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) experiment. (G and H) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-E2F-1 and anti-FOXM1 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the EZH2 promoters in qPCR. The enrichment of E2F-1 and FOXM1 on EZH2 promoter relative to IgG in HeLa cells, H3 against RPL30 used as positive control. (I and J) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-E2F-1 and anti-FOXM1 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for the indicated 4 regions in the DNMT3A promoters in qPCR. The enrichment of E2F-1 and FOXM1 on DNMT3A promoter relative to IgG in HeLa cells, H3 against RPL30 used as a positive control. **P<0.01; ns, not significant.

Journal: Oncology Reports

Article Title: Crosstalk between histone modification and DNA methylation orchestrates the epigenetic regulation of the costimulatory factors, Tim-3 and galectin-9, in cervical cancer

doi: 10.3892/or.2019.7388

Figure Lengend Snippet: HPV18 oncoprotein E6/E7 alters the expression EZH2 and H3K27me3 in cervical cancer cells. (A and B) C33A cells were transfected with 18E6/E7/E6/7 lentiviral vector and HeLa cells were transfected with 18E6/E7/E6/7 specific siRNAs and western blot analysis was then used to detect EZH2 and H3K27me3 expression. (C) EZH2 and H3K27me3 expression in HeLa and C33A cells detected by western blot analysis. (D) Western blot analysis of the HPV18 E6/E7 protein level in HeLa cells following transfection with siRNAs against EZH2. (E) Co-IP assay was used to analyzed the incorporation between EZH2 and HPV18 E6/E7. (F) Schematic representation of the 4 regions of the EZH2 promoter amplified in the chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) experiment. (G and H) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-E2F-1 and anti-FOXM1 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the EZH2 promoters in qPCR. The enrichment of E2F-1 and FOXM1 on EZH2 promoter relative to IgG in HeLa cells, H3 against RPL30 used as positive control. (I and J) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-E2F-1 and anti-FOXM1 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for the indicated 4 regions in the DNMT3A promoters in qPCR. The enrichment of E2F-1 and FOXM1 on DNMT3A promoter relative to IgG in HeLa cells, H3 against RPL30 used as a positive control. **P<0.01; ns, not significant.

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot, Co-Immunoprecipitation Assay, Amplification, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Positive Control

The HPV18 pathway regulates Tim-3/galectin-9 through EZH2-H2K27me3-DNMT3A. HPV18 E6 and E7 regulates EZH2 through direct binding of FOXM1 and E2F-1 the EZH2 promoter in order to activate EZH2 expression; in turn, H3K27me3 is upregulated. EZH2 and H3K27me3 directly interact with the DNMT3A promoter to downregulate its expression. When DNMT3A is decreased, the HAVCR2/LGALS9 promoter region is also demethylated and Tim-3/galectin-9 expression is upregulated.

Journal: Oncology Reports

Article Title: Crosstalk between histone modification and DNA methylation orchestrates the epigenetic regulation of the costimulatory factors, Tim-3 and galectin-9, in cervical cancer

doi: 10.3892/or.2019.7388

Figure Lengend Snippet: The HPV18 pathway regulates Tim-3/galectin-9 through EZH2-H2K27me3-DNMT3A. HPV18 E6 and E7 regulates EZH2 through direct binding of FOXM1 and E2F-1 the EZH2 promoter in order to activate EZH2 expression; in turn, H3K27me3 is upregulated. EZH2 and H3K27me3 directly interact with the DNMT3A promoter to downregulate its expression. When DNMT3A is decreased, the HAVCR2/LGALS9 promoter region is also demethylated and Tim-3/galectin-9 expression is upregulated.

Techniques: Binding Assay, Expressing

(A) Immunoblot analysis of DNMTs and MBD2 protein. Equal amounts of protein (80 μg) from whole-cell lysates of the control MCF-7 and MDA-MB-231 cells, treated as in Fig. 1, were separated by SDS-PAGE and subjected to Western blot analysis with specific antibodies against DNMT1, DNMT3a, DNMT3b and MBD2. Equivalence of protein loading was demonstrated by immunoblotting with anti-GAPDH antibody. (B) Relative protein levels (mean± SD) of DNMTs (n = 3). (C) Relative protein levels (mean± SD) of MBD2 (n = 3). (D) ChIP analysis of association of DNMT1 and ERα promoter. Cross-linked chromatin prepared from ER-positive MCF-7 and ER-negative MDA-MB-231 human breast cancer cells was immunoprecipitated with antibodies DNMT1. MDA-MB-231 cells treated as in Fig. 1. The immunoprecipitates were subjected to PCR analysis. Aliquots of chromatin taken before immunoprecipitation were used as input controls (n = 3). # P <0.05 compared with control cells. * P <0.05 compared with cells exposed to 1μM arsenic trioxide.

Journal: PLoS ONE

Article Title: Arsenic Induces Functional Re-Expression of Estrogen Receptor α by Demethylation of DNA in Estrogen Receptor-Negative Human Breast Cancer

doi: 10.1371/journal.pone.0035957

Figure Lengend Snippet: (A) Immunoblot analysis of DNMTs and MBD2 protein. Equal amounts of protein (80 μg) from whole-cell lysates of the control MCF-7 and MDA-MB-231 cells, treated as in Fig. 1, were separated by SDS-PAGE and subjected to Western blot analysis with specific antibodies against DNMT1, DNMT3a, DNMT3b and MBD2. Equivalence of protein loading was demonstrated by immunoblotting with anti-GAPDH antibody. (B) Relative protein levels (mean± SD) of DNMTs (n = 3). (C) Relative protein levels (mean± SD) of MBD2 (n = 3). (D) ChIP analysis of association of DNMT1 and ERα promoter. Cross-linked chromatin prepared from ER-positive MCF-7 and ER-negative MDA-MB-231 human breast cancer cells was immunoprecipitated with antibodies DNMT1. MDA-MB-231 cells treated as in Fig. 1. The immunoprecipitates were subjected to PCR analysis. Aliquots of chromatin taken before immunoprecipitation were used as input controls (n = 3). # P <0.05 compared with control cells. * P <0.05 compared with cells exposed to 1μM arsenic trioxide.

Article Snippet: Rabbit polyclonal antibodies against ERα, β-actin, and GAPDH were obtained from NeoMarkers (Fremont, CA, USA), and the mouse monoclonal antibodies against DNMT1,DNMT3a and DNMT3b was obtained from Abnova (Taipei, Taiwan).

Techniques: Western Blot, Control, SDS Page, Immunoprecipitation

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1) Product Images from "Up-regulation of DNA-methyltransferase 3A expression is associated with hypomethylation of intron 25 in human testicular germ cell tumors."

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