Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537)

Overview

  • Product name
    Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade
    See all Dnmt1 primary antibodies
  • Description
    Mouse monoclonal [60B1220.1] to Dnmt1 - ChIP Grade
  • Specificity
    This antibody detects a ~180 kDa protein, corresponding to the apparent molecular mass of Dnmt1 on SDS-PAGE immunoblots in samples of human and mouse origin. Immunogen itself has been shown to be toxic.
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, ChIP, Flow Cyt, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide:

    EKDDREDKENAFKR

    , corresponding to amino acids 637-650 of Human Dnmt1

  • Positive control
    • Human kidney (IHC), mouse ES or NIH 3T3 cell lysate (WB)
  • General notes
    For maximum product recovery, centrifuge the product vial before removing cap. Shipped at on gel packs.

Properties

Applications

Our Abpromise guarantee covers the use of ab13537 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
IHC-P Use a concentration of 1 - 2 µg/ml. DNMT1 high levels are toxic, as a result it may be tough to find sometimes. Signal amplification might be needed.
IHC-Fr Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
IP Use 2µg for 106 cells.

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.
  • 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.
  • Information by UniProt
  • Database links
  • 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

Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade images

  • Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537) at 1/1000 dilution + 0.1ug mouse recombinant protein. Additional lower band is probably degraded or in other ways modified Dnmt1.

    Predicted band size : 183 kDa
  • ab13537 at 1/10000 dilution staining Dnmt1 in mouse brain tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin-embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave with two, 5 minutes incubation intervals in citrate buffer. An antibody amplifier™ was used for staining. A HRP-conjugated anti-mouse secondary was used at 1/10 dilution.

  • Overlay histogram showing HeLa cells stained with ab13537 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13537, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • All lanes : Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537) at 1 µg/ml

    Lane 1 : HCT116 whole cell lysate
    Lane 2 : HCT116 DNMT1 KO whole cell lysate
    Lane 3 : HCT116 DNMT3b KO whole cell lysate

    Secondary
    HRP conjugated Sheep anti-mouse IgG
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 183 kDa


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537)

This product has been referenced in:
  • Chen S  et al. Histone H2B monoubiquitination is a critical epigenetic switch for the regulation of autophagy. Nucleic Acids Res 45:1144-1158 (2017). WB ; Human . Read more (PubMed: 28180298) »
  • Zhang D  et al. Non-CpG methylation by DNMT3B facilitates REST binding and gene silencing in developing mouse hearts. Nucleic Acids Res 45:3102-3115 (2017). WB ; Mouse . Read more (PubMed: 27956497) »

See all 37 Publications for this product

Product Wall

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (type I endometrial cancer tissues)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate buffer/trypsin
Permeabilization
No
Specification
type I endometrial cancer tissues
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 31 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (esophagus squamous cell)
Gel Running Conditions
Reduced Denaturing (6%gel)
Loading amount
20 µg
Specification
esophagus squamous cell
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3µg/mL · Temperature: 25°C
Username

Mr. JUNGHO YOON

Verified customer

Submitted Aug 16 2017

Application
ChIP
Sample
Human Cell lysate - whole cell (Breast cancer line)
Specification
Breast cancer line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Username

Abcam user community

Verified customer

Submitted Mar 14 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse colon cells)
Gel Running Conditions
Reduced Denaturing (12.5%)
Loading amount
100000 cells
Specification
Mouse colon cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jan 26 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Ferret Tissue lysate - whole (organotypic slices of whole brain tissue)
Gel Running Conditions
Reduced Denaturing (3-8% Tris-acetate gel% Tris-acetate gel)
Loading amount
40 µg
Specification
organotypic slices of whole brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Francis Djankpa

Verified customer

Submitted Apr 29 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Fibroblast)
Specification
Fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 30 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 17 2015

Application
ChIP
Sample
Mouse Cell lysate - whole cell (mouse)
Negative control
IgG mouse antibody
Specification
mouse
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1%formaldehyde
Positive control
none
Username

Abcam user community

Verified customer

Submitted Apr 16 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (Cardiomyocytes)
Specification
Cardiomyocytes
Treatment
Glucose, 100mM
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 15 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient gel)
Sample
Human Cell lysate - whole cell (HCT116)
Specification
HCT116
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 03 2014

1-10 of 41 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up