Anti-KDM6B / JMJD3 antibody (ab38113)

Rabbit polyclonal KDM6B / JMJD3 antibody. Validated in IHC, ChIP, ICC/IF and tested in Mouse, Human. Cited in 15 publication(s). Independently reviewed in 8 review(s).

Overview

  • Product name

    Anti-KDM6B / JMJD3 antibody
    See all KDM6B / JMJD3 primary antibodies
  • Description

    Rabbit polyclonal to KDM6B / JMJD3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human KDM6B/ JMJD3 aa 1-100 conjugated to keyhole limpet haemocyanin.
    Database link: O15054
    (Peptide available as ab38112)

  • Positive control

    • IHC-P: FFPE Human Normal Liver tissue sections. ICC/IF: HeLa cell line

Properties

Applications

Our Abpromise guarantee covers the use of ab38113 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 Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
ChIP Use at an assay dependent concentration. PubMed: 23932971

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

  • Chromatin was prepared from mouse hippocampus. Crosslinking (X-ChIP) was performed for 15 minutes with 1% formaldehyde. ChIP was performed with ab38113 at 1/50 dilution (black) or normal rabbit IgG (white). The positive control was Beta-actin. The negative control was HoxA1 (a polycomb repressed gene in hippocampus).

    See Abreview

  • ab38113 stained in HeLa cells. Cells were fixed with 4% paraformaldehyde (10 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab38113 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • IHC image of KDM6B/JMJD3 staining in a section of formalin-fixed paraffin-embedded normal human liver* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab38113, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

References

This product has been referenced in:

  • Li SH  et al. JMJD3 expression is an independent prognosticator in patients with esophageal squamous cell carcinoma. Surgery N/A:N/A (2019). Read more (PubMed: 30678869) »
  • Tottone L  et al. Histone Modifications Drive Aberrant Notch3 Expression/Activity and Growth in T-ALL. Front Oncol 9:198 (2019). Read more (PubMed: 31001470) »
See all 20 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Panc1 cell line)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
Panc1 cell line
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 14 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Mr. Bryan Niedenberger

Verified customer

Submitted Jan 31 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (L929)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
L929
Blocking step
Odyssey Blocking Buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 18°C

Abcam user community

Verified customer

Submitted Aug 30 2017

Application
ChIP
Detection step
Real-time PCR
Sample
Mouse Tissue lysate - nuclear (Hippocampus)
Specification
Hippocampus
Negative control
HoxA1, a Polycomb repressed gene in hippocampus, and normal Rabbit IgG
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehide 1%
Positive control
b-actin

Dr. Ernest Palomer Vila

Verified customer

Submitted Nov 11 2014

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing
Sample
Chicken Tissue lysate - whole (Brain)
Specification
Brain
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Ms. Ying Jiang

Verified customer

Submitted Aug 07 2013

Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

Here ab84200 and ab85392 are tested and guaranteed for WB in human samples. ab85392 also in mouse samples.

ab38113 is a fast track antibody and not tested and guaranteed in WB yet. Please see the terms and conditions for testing fast track antibodies on the datasheet of ab38113:

https://www.abcam.com/index.html?datasheet=38113 (or use the following: https://www.abcam.com/index.html?datasheet=38113).


If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More

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
Western blot
Sample
Human Cell lysate - whole cell (HeLa, HEK293, Jurkat, HL-60)
Loading amount
20 µg
Specification
HeLa, HEK293, Jurkat, HL-60
Gel Running Conditions
Reduced Denaturing (Pierce precise protein gel 4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Mr. Antibody Solutions

Verified customer

Submitted Feb 22 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Mar 16 2009

Application
Western blot
Sample
Human Cell lysate - whole cell (keratinocyte)
Loading amount
100 µg
Specification
keratinocyte
Gel Running Conditions
Reduced Denaturing (8%)
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 Jun 18 2008

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