Overview

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab36938 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 Use a concentration of 1 µg/ml. Predicted molecular weight: 154 kDa.Can be blocked with Human KDM6A / UTX peptide (ab36937).
ICC/IF Use a concentration of 1 µg/ml.
ChIP Use at an assay dependent concentration. PubMed: 26306033

Target

  • Function
    Histone demethylase that specifically demethylates 'Lys-27' of histone H3, thereby playing a central role in histone code. Demethylates trimethylated and dimethylated but not monomethylated H3 'Lys-27'. Plays a central role in regulation of posterior development, by regulating HOX gene expression. Demethylation of 'Lys-27' of histone H3 is concomitent with methylation of 'Lys-4' of histone H3, and regulates the recruitment of the PRC1 complex and monoubiquitination of histone H2A.
  • Sequence similarities
    Belongs to the UTX family.
    Contains 1 JmjC domain.
    Contains 8 TPR repeats.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • bA386N14.2 (ubiquitously transcribed X chromosome tetratricopeptide repeat protein (UTX) antibody
    • bA386N14.2 antibody
    • bA386N14.2 ubiquitously transcribed X chromosome tetratricopeptide repeat protein UTX antibody
    • DKFZp686A03225 antibody
    • Histone demethylase UTX antibody
    • KABUK2 antibody
    • Kdm6a antibody
    • KDM6A_HUMAN antibody
    • Lysine (K) specific demethylase 6A antibody
    • Lysine K specific demethylase 6A antibody
    • Lysine specific demethylase 6A ubiquitously transcribed tetratricopeptide repeat gene X chromosome antibody
    • Lysine-specific demethylase 6A antibody
    • MGC141941 antibody
    • Ubiquitously transcribed tetratricopeptide repeat X chromosome antibody
    • Ubiquitously transcribed tetratricopeptide repeat protein X linked antibody
    • Ubiquitously transcribed TPR gene on the X chromosome antibody
    • Ubiquitously transcribed TPR protein on the X chromosome antibody
    • Ubiquitously transcribed X chromosome tetratricopeptide repeat protein antibody
    • Ubiquitously-transcribed TPR protein on the X chromosome antibody
    • Ubiquitously-transcribed X chromosome tetratricopeptide repeat protein antibody
    • UTX antibody
    see all

Images

  • Lanes 1 & 3: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2 & 4: KDM6A knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Green - ab36938 observed at 154 kDa.
    Lanes 3 - 4: Red - loading control, ab18058, observed at 130 kDa.

    ab36938 was shown to recognize KDM6A in wild-type HAP1 cells as signal was lost at the expected MW in KDM6A knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and KDM6A knockout samples were subjected to SDS-PAGE. Ab36938 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 0.7 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-KDM6A / UTX antibody (ab36938) at 1 µg/ml

    Lane 1 : Whole NIH3T3 cell lysate (no shRNA)
    Lane 2 : Whole NIH3T3 cell lysate (Control shRNA)
    Lane 3 : Whole NIH3T3 cell lysate (Utx shRNA)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated Goat anti-Rabbit polyclonal

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 154 kDa
    Observed band size: 148 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Lane 3 is NIH3T3 cells transduced with lentivirus expressing shRNA for mouse Utx.

    5% milk was used as the blocking agent (incubated for 1 hour at 25°C).

    ab36938 was diluted with TBS+0.1% Tween 20 and incubated for 1 hour at 25°C. 

    See Abreview

  • All lanes : Anti-KDM6A / UTX antibody (ab36938) at 1 µg/ml

    Lane 1 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)
    Lane 2 : Testis (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG -H&L- Pre Adsorbed (HRP)
    at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 154 kDa
    Observed band size: 154,157 kDa why is the actual band size different from the predicted?
    Additional bands at: 80 kDa. We are unsure as to the identity of these extra bands.



    This Fast-Track antibody is not yet fully characterised. This image represents inconclusive preliminary data.
  • ICC/IF image of ab36938 stained MCF7 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab36938, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa cells fixed in 4% PFA at 1ug/ml and HeLa and MCF7 100% methanol fixed cells at 1ug/ml. However, this Fast-Track antibody is not yet fully characterised. This image represents inconclusive preliminary data.

References

This product has been referenced in:
  • Denisenko O  et al. Chromatin changes trigger laminin genes dysregulation in aging kidneys. Aging (Albany NY) 10:1133-1145 (2018). Read more (PubMed: 29846172) »
  • Morozov VM  et al. Inhibitor of H3K27 demethylase JMJD3/UTX GSK-J4 is a potential therapeutic option for castration resistant prostate cancer. Oncotarget 8:62131-62142 (2017). Read more (PubMed: 28977932) »
See all 19 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (7,5 %, tank blot to PVDF)
Sample
Human Cell lysate - whole cell (U2OS cell line)
Specification
U2OS cell line
Treatment
-/+ transfection with full length UTX expression plasmid
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jun 20 2013

Answer

Thank you for your reply and for confirming the steps of your protocol.


Since these antibodies are Fast Track antibodies, they are only guaranteed to work in ELISA using the peptide immunogen. These are not tested in-house, and therefore not guaranteed to work, in WB with lysates. In addition, 2 of the antibodies, ab36938 and ab38113 were purchased in 2008 so it may be that these antibodies are no longer viable.


https://www.abcam.com/index.html?pageconfig=abpromise


The protocol looks okay, but since you're not seeing the bands at the proper MW, you may want to increase the concentration of both the primary and secondary antibodies.


You can also increase the amount of protein loaded. I see that you loaded 2 x 10^5 cells/lane, but we recommend at least 30 ug.


We do not know if these protocol suggestions will help with the antibodies since we haven't validated them for usein WB so we don't have internal protocols to reference. However, I hope this information helps. Since these Fast Track antibodies aren't covered by the Abpromise, it is best to try to augment the protocol to optimize the results.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain Section)
Specification
Brain Section
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric Acid
Permeabilization
Yes - 0.1% Tween20
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Oct 08 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Pig Cell lysate - whole cell (Heart, Liver and Kidney)
Loading amount
25 µg
Specification
Heart, Liver and Kidney
Gel Running Conditions
Reduced Denaturing (NuPAGE BisTris 10% gel (invitrogen))
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 0% · Temperature: 22°C

Dr. Vanessa Hall

Verified customer

Submitted Aug 20 2010

Application
Western blot
Sample
Mouse Cell lysate - nuclear (NIH3T3 cell lysate)
Loading amount
20 µg
Specification
NIH3T3 cell lysate
Gel Running Conditions
Reduced Denaturing
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 Dec 11 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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