DNMT1 Mouse Monoclonal Antibody [A3A9]
Catalog# EM1901-84
DNMT1 Mouse Monoclonal Antibody [A3A9]
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WB
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IHC-P
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FC
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Human
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unconjugated
Overview
Product Name
DNMT1 Mouse Monoclonal Antibody [A3A9]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Synthetic peptide within Human Dnmt1 aa 500-600 / 1616.
Species Reactivity
Human
Validated Applications
WB, IHC-P, FC
Molecular Weight
183 kDa
Positive Control
Daudi cell lysate, 293 cell lysate, human tonsil tissue, JAR.
Conjugation
unconjugated
Clone Number
A3A9
RRID
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
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WB
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1:500-1:1,000
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
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
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
ExpandImages
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Western blot analysis of DNMT1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-84, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Daudi cell lysate
Lane 2: 293 cell lysate -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-DNMT1 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-84, 1/50) for 30 minutes 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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Flow cytometric analysis of DNMT1 was done on JAR cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-84, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"