Overview

  • Product name
    Anti-KDM5C / Jarid1C / SMCX antibody
    See all KDM5C / Jarid1C / SMCX primary antibodies
  • Description
    Rabbit polyclonal to KDM5C / Jarid1C / SMCX
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ChIP, IHC-FoFr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1500 to the C-terminus of Human Jarid1C/ SMCX.

    Read Abcam's proprietary immunogen policy (Peptide available as ab35501.)

  • Positive control
    • This antibody gave a positive signal in HEK 293 (Human embryonic kidney cell line), and Y79 (Human retinoblastoma cell line) Whole Cell Lysates. This antibody gave a positive signal in the following Methanol fixed cell lines:HeLa, Hek293, HepG2, and MCF-7. This antibody gave a positive signal in the following Formaldehyde fixed cell lines:HepG2, and MCF-7.

Properties

Applications

Our Abpromise guarantee covers the use of ab34718 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
ChIP Use at an assay dependent concentration. PubMed: 19136938
IHC-FoFr Use at an assay dependent concentration.
WB 1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 176 kDa).

Abcam recommends using milk as the blocking agent.

ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 'Lys-9', H3 'Lys-27', H3 'Lys-36', H3 'Lys-79' or H4 'Lys-20'. Demethylates trimethylated and dimethylated but not monomethylated H3 'Lys-4'. Participates in transcriptional repression of neuronal genes by recruiting histone deacetylases and REST at neuron-restrictive silencer elements.
  • Tissue specificity
    Expressed in all tissues examined. Highest levels found in brain and skeletal muscle.
  • Involvement in disease
    Defects in KDM5C are the cause of mental retardation syndromic X-linked JARID1C-related (MRXSJ) [MIM:300534]. MRXSJ is characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. MRXSJ patients manifest mental retardation associated with variable features such as slowly progressive spastic paraplegia, seizures, facial dysmorphism.
  • Sequence similarities
    Belongs to the JARID1 histone demethylase family.
    Contains 1 ARID domain.
    Contains 1 JmjC domain.
    Contains 1 JmjN domain.
    Contains 2 PHD-type zinc fingers.
  • Domain
    The first PHD-type zinc finger domain recognizes and binds H3-K9Me3.
    Both the JmjC domain and the JmjN domain are required for enzymatic activity.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • DXS1272E antibody
    • Histone demethylase JARID1C antibody
    • JARID1C antibody
    • JmjC domain containing protein SMCX antibody
    • Jumonji AT rich interactive domain 1C antibody
    • Jumonji, AT rich interactive domain 1C (RBP2 like) antibody
    • Jumonji/ARID domain-containing protein 1C antibody
    • KDM5C antibody
    • KDM5C_HUMAN antibody
    • Lysine (K) specific demethylase 5C antibody
    • Lysine-specific demethylase 5C antibody
    • MRX13 antibody
    • MRXJ antibody
    • MRXSCJ antibody
    • MRXSJ antibody
    • Protein SmcX antibody
    • Protein Xe169 antibody
    • rbp2 like protein antibody
    • Selected cDNA on X antibody
    • SMCX antibody
    • Smcx homolog X chromosome antibody
    • SmcX protein antibody
    • Smcy homolog X linked antibody
    • XE169 antibody
    • Xe169 protein antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: KDM5C knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HEK293 whole cell lysate (20 µg)
    Lane 4: U2OS whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab34718 observed at 175 kDa. Red - loading control, ab18058, observed at 120 kDa.

    ab34718 was shown to specifically recognize KDM5C in wild-type HAP1 cells as signal was lost at the expected MW in KDM5C knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and KDM5C knockout samples were subjected to SDS-PAGE. Ab34718 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20,000 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 1/20,000 dilution for 1 hour at room temperature before imaging.

  • PFA-fixed, 0.5% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for KDM5C / Jarid1C / SMCX (green) using ab34718 at 1/200 dilution in ICC/IF. Counter-stained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • ab34718 stained in Hela cells. Cells were fixed with 100% methanol (5min) 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 ab34718 at 5µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 µg/ml 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.

  • All lanes : Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1 µg/ml

    Lane 1 : HEK293 (Human) Whole Cell Lysate
    Lane 2 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 176 kDa
    Observed band size: 180 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 45 kDa, 58 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    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 3% Milk before being incubated with ab34718 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1/250 dilution + HEK293 whole cell lysate (ab7902) at 20 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 176 kDa
    Observed band size: 176 kDa

References

This product has been referenced in:
  • Fujita T  et al. Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP). Sci Rep 3:3171 (2013). ICC/IF ; Human . Read more (PubMed: 24201379) »
  • Liang Z  et al. Proliferating cell nuclear antigen is required for loading of the SMCX/KMD5C histone demethylase onto chromatin. Epigenetics Chromatin 4:18 (2011). Read more (PubMed: 21996408) »
See all 4 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

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

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 27 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (different cell lines)
Gel Running Conditions
Reduced Denaturing (7,5% gel)
Loading amount
50 µg
Specification
different cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 29 2013

Answer

Thank you for contacting us.

I have checked the homology between KDM5D and KDM5C, and for the immunogen region of ab34718, the homology is poor and the antibody would not cross react.

Unfortunately, we do not carry antibodies that were raised against a KDM5D sequence itself. YOu might want to check http://www.biocompare.com for such antibodies.

I hope this information is of some helpto you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (brain-perfused frozen sections)
Specification
brain-perfused frozen sections
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 30 2012

Answer

Thank you for contacting us.

The abreview (xxx) is in dormant status as it was not edited for over 28 days. I have just put it back into editing status and you should be able to see it. Lots of information is still missing which would need to be filled in before submitting it.

Since this review is for mouse and ICC, another review for the same antibody and same species/application cannot be submitted. Thus, please complete the current review. Thank you so much!

Once the review is submitted, the discount code will be active and can be used on an order.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for your email.
I am glad to hear that the antibody has worked successfully in the untested species.

Here iswhat you need to do in order to activate your discount code:


The code will give you 1 free PRIMARY ANTIBODY before the expiration date (March 2, 2012). To redeem this offer, please submit an Abreview for mouse/ICC and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

The immunogen of the antibody is available ab35501.
https://www.abcam.com/jarid1c-smcx-peptide-ab35501.html
The itemis available to order. Please contact customer serviceif you would like to place an order for this product and we would estimate that we would be able to supply this within 4-6 weeks from the date of the order.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Y79 retinoblastoma cell line)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
Y79 retinoblastoma cell line
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Sep 12 2008

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

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