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RecombinantRabMAb

Recombinant Anti-Dnmt3L antibody [EPR18774] (ab194094)

Reviews (1) Submit a question References (3)
Western blot - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Immunoprecipitation - Anti-Dnmt3L antibody [EPR18774] (ab194094)
  • Anti-Dnmt3L antibody [EPR18774] (ab194094)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18774] to Dnmt3L
  • Suitable for: IHC-P, WB, IP, ICC/IF
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Dnmt3L antibody [EPR18774]
    See all Dnmt3L primary antibodies

  • Description

    Rabbit monoclonal [EPR18774] to Dnmt3L

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, IP, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: mESC, F9 and NCCIT whole cell lysates. IHC-P: Human seminoma and yolk sac tumor tissues. ICC/IF: NCCIT cells. IP: F9 whole cell lysate.

  • 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.

Properties

  • 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

    EPR18774

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab194094 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
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested using Human tissues only
WB
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).

Predicted and actual band for mouse = 48kDa.Predicted and actual band for human = 44kDa.
IP
1/50.
ICC/IF
1/250.
Notes
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested using Human tissues only
WB
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).

Predicted and actual band for mouse = 48kDa.Predicted and actual band for human = 44kDa.
IP
1/50.
ICC/IF
1/250.

Target

  • Function

    Catalytically inactive regulatory factor of DNA methyltransferases. It is essential for the function of DNMT3A and DNMT3B. Activates DNMT3A and DNMT3B by binding to their catalytic domain. Accelerates the binding of DNA and AdoMet to the methyltransferases and dissociates from the complex after DNA binding to the methyltransferases. Recognizes unmethylated histone H3 lysine 4 (H3K4) and induces de novo DNA methylation by recruitment or activation of DNMT3.

  • Tissue specificity

    Expressed at low levels in several tissues including testis, ovary, and thymus.

  • Sequence similarities

    Belongs to the C5-methyltransferase family.
    Contains 1 ADD-type zinc finger.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Cytosine 5 methyltransferase 3 like protein antibody
      • DNA (cytosine 5 ) methyltransferase 3 like antibody
      • DNA (cytosine-5)-methyltransferase 3-like antibody
      • DNA cytosine 5 methyltransferase 3 like protein antibody
      • DNA methyltransferase 3 like protein antibody
      • DNM3L_HUMAN antibody
      • Dnmt 3L antibody
      • Dnmt3l antibody
      • Human cytosine 5 methyltransferase 3 like protein antibody
      • MGC1090 antibody
      see all

    Images

    • Western blot - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Western blot - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      All lanes : Anti-Dnmt3L antibody [EPR18774] (ab194094) at 1/1000 dilution

      Lane 1 : mESC (Mouse embryonic stem cells ) whole cell lysate
      Lane 2 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate
      Lane 3 : Rat testis lysate
      Lane 4 : NTERA-2 cl.D1 [NT2/D1] (Human malignant pluripotent embryonic carcinoma) whole cell lysate
      Lane 5 : NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate
      Lane 6 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
      Lane 7 : T-47D (Human ductal breast epithelial tumor 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: 48 kDa
      Observed band size: 44 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Observed MW: 48 kDa (mouse), 44 kDa (human).

      Exposure time: Lanes 1 & 2: 1 second; Lanes 3,4,5,6 & 7: 3 minutes. GAPDH control lanes: 30 seconds.

      1. The predicted molecular weight of human Dnmt3L is 44kDa and the predicted molecular weight of mouse Dnmt3L is 48kDa. This is consistent with what has been observed.
      2. HeLa and T-47D cells do not express Dnmt3L (PMID: 20460473).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)

      Immunohistochemical analysis of paraffin-embedded Human seminoma tissue labeling Dnmt3L with ab194094 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Mainly nuclear with weak cytoplasmic staining was found on tumor cells of Human seminoma.

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)

      Immunohistochemical analysis of paraffin-embedded Human yolk sac tumor tissue labeling Dnmt3L with ab194094 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Nuclear staining was found on the tumor cells of Human yolk sac tumor.

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Dnmt3L with ab194094 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Negative staining on the Human liver is observed.

      Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (ab194094)

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Dnmt3L with ab194094 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Negative staining on the Human liver is observed.

      Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3L antibody [EPR18774] (ab194094)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (Human pluripotent embryonic carcinoma) and HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Dnmt3L with ab194094 at 1/250 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 NCCIT cell line. Negative expression in HeLa cells (PMID: 20460473).

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab194094 at 1/250 dilution followed by ab150120 at 1/1000 dilution.

      -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

    • Immunoprecipitation - Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Immunoprecipitation - Anti-Dnmt3L antibody [EPR18774] (ab194094)

      Dnmt3L was immunoprecipitated from 1mg of F9 (Mouse embryonic testicular cancer cell line) whole cell lysate with ab194094 at 1/50 dilution.

      Western blot was performed from the immunoprecipitate using ab194094 at 1/1000 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: F9 whole cell lysate,10ug (Input).

      Lane 2: ab194094 IP in F9 whole cell lysate.

      Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab191594 in F9 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

    • Anti-Dnmt3L antibody [EPR18774] (ab194094)
      Anti-Dnmt3L antibody [EPR18774] (ab194094)

    References (3)

    Publishing research using ab194094? Please let us know so that we can cite the reference in this datasheet.

    ab194094 has been referenced in 3 publications.

    • Thomas J  et al. Running the full human developmental clock in interspecies chimeras using alternative human stem cells with expanded embryonic potential. NPJ Regen Med 6:25 (2021). PubMed: 34001907
    • Ochiai H  et al. Genome-wide kinetic properties of transcriptional bursting in mouse embryonic stem cells. Sci Adv 6:eaaz6699 (2020). PubMed: 32596448
    • Ishiuchi T  et al. Zfp281 Shapes the Transcriptome of Trophoblast Stem Cells and Is Essential for Placental Development. Cell Rep 27:1742-1754.e6 (2019). PubMed: 31067460

    Customer reviews and Q&As

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    Flow Cytometry abreview for Anti-Dnmt3L antibody [EPR18774]

     Excellent
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (K562 cell line nucleofected with DNMT3L-IRES-GFP p)
    Permeabilization
    Yes - BD Pharmingen Transcription Factor Buffer Set
    Gating Strategy
    GFP+
    Specification
    K562 cell line nucleofected with DNMT3L-IRES-GFP p
    Fixation
    BD Pharmingen Transcription Factor Buffer Set
    Read More
    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 13 2022

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