Name: Anti-Dnmt1 antibody [EPR18453]
Brand: RabMAb
SKU: ab188453
Rating: 5 (7 reviews)

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Western blot - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18453] to Dnmt1
Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

Product name

Anti-Dnmt1 antibody [EPR18453]
See all Dnmt1 primary antibodies
Description

Rabbit monoclonal [EPR18453] to Dnmt1
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293, Jurkat, HeLa, Neuro-2a and NIH/3T3 whole cell lysates. IHC-P: Human placenta, mouse stomach and rat stomach tissues. ICC/IF: Wild-type HAP1, HeLa, 293T and Neuro-2a cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18453
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Dnmt1 protein (ab198109)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab188453 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P	(1)	1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF	(2)	Use a concentration of 1 µg/ml.
WB	(3)	1/1000. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa).

Notes
Flow Cyt (Intra) Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
WB 1/1000. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa).

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.
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.
Sequence similarities

Belongs to the C5-methyltransferase family.
Contains 2 BAH domains.
Contains 1 CXXC-type zinc finger.
Domain

The N-terminal part is required for homodimerization and acts as a regulatory domain.
Post-translational
modifications

Sumoylated; sumoylation increases activity.
Cellular localization

Nucleus.
Target information above from: UniProt accession P26358 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 1786 Human
- Entrez Gene: 13433 Mouse
- Entrez Gene: 84350 Rat
- Omim: 126375 Human
- SwissProt: P26358 Human
- SwissProt: P13864 Mouse
- SwissProt: Q9Z330 Rat
- Unigene: 202672 Human
- Unigene: 128580 Mouse
- Unigene: 6955 Rat
see all
Alternative names
- 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
- DNA MTase antibody
- DNA MTase HsaI antibody
- DNMT 1 antibody
- DNMT antibody
- Dnmt1 antibody
- DNMT1_HUMAN antibody
- Dnmt1o antibody
- FLJ16293 antibody
- HSN1E antibody
- M.HsaI antibody
- MCMT antibody
- Met1 antibody
- MGC104992 antibody
- mMmul antibody
- MommeD2 antibody
see all

Western blot - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 µg)

Lanes 1 - 2: Merged signal (red and green). Green - ab188453 observed at 183 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab188453 was shown to specifically react with DNMT1 when DNMT1 knockout samples were used. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. Ab188453 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody [EPR18453] (ab188453)

ab188453 staining Dnmt1 in wild-type HAP1 cells (top panel) and DNMT1 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab188453 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling Dnmt1 with ab188453 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Neuro-2a cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab188453 at 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma)cells labeling with purified ab188453 at 1/100 dilution (10ug/ml) (Red). Cells were fixed with4% paraformaldehydeand permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody.Rabbit monoclonal IgG (Black) (ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
Western blot - Anti-Dnmt1 antibody [EPR18453] (ab188453)

All lanes : Anti-Dnmt1 antibody [EPR18453] (ab188453) at 1/1000 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Dnmt1 with ab188453 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human placenta is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Dnmt1 antibody [EPR18453] (ab188453)

All lanes : Anti-Dnmt1 antibody [EPR18453] (ab188453) at 1/200 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Dnmt1 with ab188453 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse stomach is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Dnmt1 with ab188453 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab188453 at 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Dnmt1 with ab188453 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat stomach is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody [EPR18453] (ab188453)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling Dnmt1 with ab188453 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HEK-293 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab188453 at 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Anti-Dnmt1 antibody [EPR18453] (ab188453)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab188453? Please let us know so that we can cite the reference in this datasheet.

ab188453 has been referenced in 35 publications.

Li H et al. miR-142-5p Inhibits Cell Invasion and Migration by Targeting DNMT1 in Breast Cancer. Oncol Res 28:885-897 (2022). PubMed: 34321149
Chen B et al. Regulation of paternal 5mC oxidation and H3K9me2 asymmetry by ERK1/2 in mouse zygotes. Cell Biosci 12:25 (2022). PubMed: 35255956
Guan H et al. DNA methyltransferase 1 knockdown reverses PTEN and VDR by mediating demethylation of promoter and protects against renal injuries in hepatitis B virus-associated glomerulonephritis. Cell Biosci 12:98 (2022). PubMed: 35765066
Tan Y et al. RNA interference-mediated silencing of DNA methyltransferase 1 attenuates neuropathic pain by accelerating microglia M2 polarization. BMC Neurol 22:376 (2022). PubMed: 36183073
Yu Y et al. The lncRNA KIF9-AS1 Accelerates Hepatocellular Carcinoma Growth by Recruiting DNMT1 to Promote RAI2 DNA Methylation. J Oncol 2022:3888798 (2022). PubMed: 36276278

View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - other (Brain tumor)

Gel Running Conditions

Reduced Denaturing (4-12%)

Loading amount

10 µg

Specification

Brain tumor

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ivano Mocavini

Verified customer

Submitted Feb 05 2021

Application

Western blot

Sample

Guinea pig Cell lysate - whole cell (Liver)

Gel Running Conditions

Reduced Denaturing (Stacking 4%, separating 6%)

Loading amount

50 µg

Treatment

Deficient diet

Specification

Liver

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 19 2019

Application

Western blot

Sample

Chicken Cell lysate - whole cell (Chincken B cell line DT40)

Gel Running Conditions

Reduced Denaturing (3-8% Tris-acetate)

Loading amount

100000 cells

Specification

Chincken B cell line DT40

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 22 2016

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (35)

Customer reviews and Q&As

Western blot abreview for Anti-Dnmt1 antibody [EPR18453]

Western blot abreview for Anti-Dnmt1 antibody [EPR18453]

Western blot abreview for Anti-Dnmt1 antibody [EPR18453]

Post-translational
modifications