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

Mouse monoclonal Dnmt1 antibody [60B1220.1]. Validated in WB, IP, IHC, Flow Cyt, ChIP and tested in Mouse, Rat, Human, Zebrafish. Cited in 81 publication(s). Independently reviewed in 22 review(s).

Overview

  • Product name
    Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade
    See all Dnmt1 primary antibodies
  • Description
    Mouse monoclonal [60B1220.1] to Dnmt1 - ChIP Grade
  • Host species
    Mouse
  • 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
    • IHC-P: Mouse brain and kidney medullar tissue. Flow cyt: HeLa cells. WB: HCT 116 cells; Dnmt1 recombinant protein. 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

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.

  • Mouse medullar kidney tissue stained for Dnmt1 using ab13537 at 1/1000 dilution in immunohistochemical analysis. DAB staining. Counterstained with hematoxylin.

  • 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
    All lanes : HRP conjugated Sheep anti-mouse IgG

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 183 kDa


    Exposure time: 1 minute

    See Abreview

References

This product has been referenced in:
  • Mishra M & Kowluru RA DNA Methylation-a Potential Source of Mitochondria DNA Base Mismatch in the Development of Diabetic Retinopathy. Mol Neurobiol 56:88-101 (2019). Read more (PubMed: 29679259) »
  • Jin J  et al. Methylation-associated silencing of miR-128 promotes the development of esophageal cancer by targeting COX-2 in areas with a high incidence of esophageal cancer. Int J Oncol 54:644-654 (2019). Read more (PubMed: 30535495) »
See all 101 Publications for this product

Customer reviews and Q&As

11-20 of 42 Abreviews or Q&A

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

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

Abcam user community

Verified customer

Submitted Nov 03 2014

Application
ChIP
Detection step
Real-time PCR
Sample
Rat Tissue lysate - whole (GONAD)
Specification
GONAD
Negative control
IgG
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Positive control
Input DNA

Abcam user community

Verified customer

Submitted Jul 07 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (6%)
Sample
Mouse Cell lysate - whole cell (Mouse N2A neuroblastoma)
Specification
Mouse N2A neuroblastoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 03 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (6%)
Sample
Human Cell lysate - whole cell (hek 293)
Specification
hek 293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 02 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step
Enzymatic
Sample
Rat Tissue sections (GONAD)
Specification
GONAD
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 23 2013

Application
Immunoprecipitation
Immuno-precipitation step
Protein A
Sample
Human Cell lysate - whole cell (fibroblasts)
Specification
fibroblasts
Total protein in input
100000 cells

Abcam user community

Verified customer

Submitted Aug 19 2013

Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing (12 %)
Sample
Human Cell lysate - other (fibroblasts)
Specification
fibroblasts
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 14 2013

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1208729 (ab92453).

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for your email.

We have decided to send you a replacement antibody. The antibody ab92453 is of same clone and same immunogen sequence. Could you check you and confirm?

Please note, although ab13537 and ab92453 seems same however they are developed for us by different laboratories so we can expect some variations as different preparations of the hybridoma are known to alter epitope specificity slightly.

Please also note that as of ab13537 this antibody has also not been tested in double staining where the second antibody was against same antigen. So protocol optimizations are highly recommended.

I will look forward to hearing from you soon. Please do not hesitate to contact me if you have any question.

Read More

11-20 of 42 Abreviews or Q&A

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