Overview

  • Product name
    Anti-KDM6B / JMJD3 antibody
    See all KDM6B / JMJD3 primary antibodies
  • Description
    Rabbit polyclonal to KDM6B / JMJD3
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Cow, Chimpanzee, Macaque monkey, Gorilla
  • Immunogen

    Synthetic peptide corresponding to Mouse KDM6B/ JMJD3 aa 50-150 conjugated to keyhole limpet haemocyanin.
    Database link: Q5NCY0

  • Positive control
    • This antibody gave a positive signal in both HeLa whole cell and HeLa Nuclear lysates. This antibody gave a positive result when used in the following methanol fixed cell lines: MCF-7

Properties

Applications

Our Abpromise guarantee covers the use of ab169197 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 177 kDa (predicted molecular weight: 177 kDa).

Target

  • Function
    Histone demethylase that specifically demethylates 'Lys-27' of histone H3, thereby playing a central role in histone code. Demethylates trimethylated and dimethylated H3 'Lys-27'. Plays a central role in regulation of posterior development, by regulating HOX gene expression. Involved in inflammatory response by participating in macrophage differentiation in case of inflammation by regulating gene expression and macrophage differentiation.
  • Sequence similarities
    Belongs to the UTX family.
    Contains 1 JmjC domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Histone demethylase JMJD3 antibody
    • JmjC domain containing protein 3 antibody
    • JmjC domain-containing protein 3 antibody
    • Jumonji D3 antibody
    • Jumonji domain containing 3 antibody
    • Jumonji domain containing 3 histone lysine demethylase antibody
    • Jumonji domain containing protein 3 antibody
    • Jumonji domain-containing protein 3 antibody
    • Kdm6b antibody
    • KDM6B_HUMAN antibody
    • KIAA0346 antibody
    • Lysine demethylase 6B antibody
    • Lysine K specific demethylase 6B antibody
    • Lysine specific demethylase 6B antibody
    • Lysine-specific demethylase 6B antibody
    see all

Images



  • Predicted band size : 177 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: KDM6B knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lanes 1 - 3: Merged signal (red and green). Green - ab169197 observed at 120 kDa. Red - loading control, ab18058.

    ab169197 was shown to specifically react with KDM6B when KDM6B knockout samples were used. Wild-type and KDM6B knockout samples were subjected to SDS-PAGE.  Ab169197 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • ICC/IF image of ab169197 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab169197 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-KDM6B / JMJD3 antibody (ab169197) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 177 kDa
    Observed band size : 177 kDa


    Exposure time : 4 minutes

    Due to the cellular localisation of KDM6B / JMJD3 an increase in expression is expected within the Nuclear fraction.

    This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab169197 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

This product has been referenced in:
  • Li Q  et al. KDM6B induces epithelial-mesenchymal transition and enhances clear cell renal cell carcinoma metastasis through the activation of SLUG. Int J Clin Exp Pathol 8:6334-44 (2015). Read more (PubMed: 26261509) »

See 1 Publication for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Western blot
Sample
Rat Tissue lysate - whole (spinal cord)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
70 µg
Specification
spinal cord
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Sep 15 2015

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