Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [EPR18774] to Dnmt3L

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Mouse
    WB
    Human
    See all applications and species data

  • 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

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)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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