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DNMT1 Mouse Monoclonal Antibody [A3A8]

Usd: 350

Specification

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Catalog# EM1901-83

DNMT1 Mouse Monoclonal Antibody [A3A8]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

DNMT1 Mouse Monoclonal Antibody [A3A8]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human Dnmt1 aa 500-600 / 1616.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 183 kDa

Positive Control

HeLa cell lysate, 293T cell lysate, HepG2 cell lysate, Jurkat cell lysate, SH-SY5Y cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, mouse ovary tissue lysate, mouse testis tissue lysate, rat ovary tissue lysate, rat testis tissue lysate, HeLa, human testis tissue, human tonsil tissue, mouse ovary tissue, mouse testis tissue, rat ovary tissue, rat testis tissue.

Conjugation

unconjugated

Clone Number

A3A8

RRID

AB_3068928

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:4,000

  • IHC-P

  • 1:200-1:1,000

  • IF-Cell

  • 1:400

Target

Function

Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9. Probably forms a corepressor complex required for activated KRAS-mediated promoter hypermethylation and transcriptional silencing of tumor suppressor genes (TSGs) or other tumor-related genes in colorectal cancer (CRC) cells. Also required to maintain a transcriptionally repressive state of genes in undifferentiated embryonic stem cells (ESCs). Associates at promoter regions of tumor suppressor genes (TSGs) leading to their gene silencing. Promotes tumor growth.

Background References

1. Li T. et. al. Structural and mechanistic insights into UHRF1-mediated DNMT1 activation in the maintenance DNA methylation. Nucleic Acids Res. 2018 Apr 6;46(6):3218-3231.

2. Du WW. et. al. A circular RNA circ-DNMT1 enhances breast cancer progression by activating autophagy. Oncogene. 2018 Nov;37(44):5829-5842.

Sequence Similarity

Belongs to the class I-like SAM-binding methyltransferase superfamily. C5-methyltransferase family.

Tissue Specificity

Ubiquitous; highly expressed in fetal tissues, heart, kidney, placenta, peripheral blood mononuclear cells, and expressed at lower levels in spleen, lung, brain, small intestine, colon, liver, and skeletal muscle. Isoform 2 is less expressed than isoform 1.

Post-translational Modification

Sumoylated; sumoylation increases activity.; Acetylation on multiple lysines, mainly by KAT2B/PCAF, regulates cell cycle G(2)/M transition. Deacetylation of Lys-1349 and Lys-1415 by SIRT1 increases methyltransferase activity.; Phosphorylation of Ser-154 by CDKs is important for enzymatic activity and protein stability. Phosphorylation of Ser-143 by AKT1 prevents methylation by SETD7 therebye increasing DNMT1 stability.; Methylation at Lys-142 by SETD7 promotes DNMT1 proteasomal degradation.; Ubiquitinated by UHRF1; interaction with USP7 counteracts ubiquitination by UHRF1 by promoting deubiquitination and preventing degradation by the proteasome.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P26358 Human

SwissProt: P13864 Mouse

SwissProt: Q9Z330 Rat

Synonyms

ADCADN antibody

AIM antibody

CXXC finger protein 9 antibody

CXXC-type zinc finger protein 9 antibody

CXXC9 antibody

DNA (cytosine 5 ) methyltransferase 1 antibody

DNA (cytosine-5)-methyltransferase 1 antibody

DNA methyltransferase 1 antibody

DNA methyltransferase HsaI antibody

DNA methyltransferase M.HsaI. antibody

Expand

Images

  • Western blot analysis of DNMT1 on different lysates with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/4,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: 293T cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: SH-SY5Y cell lysate<br />Lane 6: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 183 kDa<br />Observed band size: 183 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/4,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of DNMT1 on different lysates with Mouse anti-DNMT1 antibody (EM1901-83) at 1/4,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: 293T cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: SH-SY5Y cell lysate
    Lane 6: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 183 kDa
    Observed band size: 183 kDa

    Exposure time: 3 minutes; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-83) at 1/4,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of DNMT1 on different lysates with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: Mouse ovary tissue lysate (40 µg/Lane)<br />Lane 4: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 5: Rat ovary tissue lysate (40 µg/Lane)<br />Lane 6: Rat testis tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 183 kDa<br />Observed band size: 183 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of DNMT1 on different lysates with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

    Lane 1: 293T cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: Mouse ovary tissue lysate (40 µg/Lane)
    Lane 4: Mouse testis tissue lysate (40 µg/Lane)
    Lane 5: Rat ovary tissue lysate (40 µg/Lane)
    Lane 6: Rat testis tissue lysate (40 µg/Lane)

    Predicted band size: 183 kDa
    Observed band size: 183 kDa

    Exposure time: 3 minutes; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-83) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling DNMT1 with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/400 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/400 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling DNMT1 with Mouse anti-DNMT1 antibody (EM1901-83) at 1/400 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-DNMT1 antibody (EM1901-83) at 1/400 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse ovary tissue with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse ovary tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat ovary tissue with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat ovary tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-DNMT1 antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-83" style="font-weight: bold;text-decoration: underline;">EM1901-83</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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Citation