Overview

  • Product name

    Anti-KDM4B / JMJD2B antibody [EPR18603]
    See all KDM4B / JMJD2B primary antibodies
  • Description

    Rabbit monoclonal [EPR18603] to KDM4B / JMJD2B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human KDM4B/ JMJD2B aa 100-200. The exact sequence is proprietary.
    Database link: O94953

  • Positive control

    • WB: T-47D, NIH/3T3, SW480, HEK-293, LNCaP, HCT 116, LLC, C6, RAW 264.7, PC-12 whole cell lysates; mouse and rat testis lysates. IHC-P: Human colon, mouse stomach and rat kidney tissues. ICC/IF: NIH/3T3 and T-47D cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab191434 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
WB 1/5000. Detects a band of approximately 150 kDa (predicted molecular weight: 122 kDa).
IHC-P 1/125. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/1000.

Target

  • Function

    Histone demethylase that specifically demethylates 'Lys-9' of histone H3, thereby playing a role in histone code. Does not demethylate histone H3 'Lys-4', H3 'Lys-27', H3 'Lys-36' nor H4 'Lys-20'. Only able to demethylate trimethylated H3 'Lys-9', with a weaker activity than KDM4A, KDM4C and KDM4D. Demethylation of Lys residue generates formaldehyde and succinate.
  • Sequence similarities

    Belongs to the JHDM3 histone demethylase family.
    Contains 1 JmjC domain.
    Contains 1 JmjN domain.
    Contains 2 PHD-type zinc fingers.
    Contains 2 Tudor domains.
  • Domain

    The 2 Tudor domains recognize and bind methylated histones. Double Tudor domain has an interdigitated structure and the unusual fold is required for its ability to bind methylated histone tails.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • JHDM3B antibody
    • JmjC domain-containing histone demethylation protein 3B antibody
    • JMJD2B antibody
    • Jumonji domain containing 2B antibody
    • Jumonji domain containing protein 2B antibody
    • Jumonji domain-containing protein 2B antibody
    • Kdm4b antibody
    • KDM4B_HUMAN antibody
    • KIAA0876 antibody
    • Lysine (K) specific demethylase 4B antibody
    • Lysine demethylase 4B antibody
    • Lysine-specific demethylase 4B antibody
    • TDRD14B antibody
    • Tudor domain containing 14B antibody
    see all

Images

  • All lanes : Anti-KDM4B / JMJD2B antibody [EPR18603] (ab191434) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cells) 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: 122 kDa
    Observed band size: 150 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-KDM4B / JMJD2B antibody [EPR18603] (ab191434) at 1/5000 dilution

    Lane 1 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysate
    Lane 3 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
    Lane 4 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 5 : LNCaP (Human prostate cancer cell line) whole cell lysate
    Lane 6 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
    Lane 7 : LLC (Mouse lung carcinoma) whole cell lysate
    Lane 8 : Mouse testis lysate
    Lane 9 : Rat testis 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: 122 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized T-47D (Human ductal breast epithelial tumor cell line) cells labeling KDM4B / JMJD2B with ab191434 at 1/1000 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 T-47D 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: ab191434 at 1/1000 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 analysis of NIH/3T3 (mouse embryo) labelling KDM4B/JMJD2B with purified ab191434 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling KDM4B / JMJD2B with ab191434 at 1/125 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human colon 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.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling KDM4B / JMJD2B with ab191434 at 1/125 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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling KDM4B / JMJD2B with ab191434 at 1/125 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat kidney 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cells) cells labeling KDM4B / JMJD2B with ab191434 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and 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: ab191434 at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

References

This product has been referenced in:

  • Ruan Z  et al. Effect of sex differences in donor foetal fibroblast on the early development and DNA methylation status of buffalo (Bubalus bubalis) nuclear transfer embryos. Reprod Domest Anim N/A:N/A (2018). Read more (PubMed: 30051521) »
  • Su Z  et al. Reader domain specificity and lysine demethylase-4 family function. Nat Commun 7:13387 (2016). IHC-Fr ; Rat . Read more (PubMed: 27841353) »
See all 2 Publications for this product

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