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  1. Link

    kdm5c--jarid1c--smcx-antibody-ab34718.pdf

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Epigenetics and Nuclear Signaling Nuclear Signaling Pathways Nuclear Receptors Co-activators/co-repressors
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Validated using a knockout cell line

Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)

  • Datasheet
Reviews (5)Q&A (4)References (7)

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Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
  • Immunocytochemistry - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
  • Immunocytochemistry/ Immunofluorescence - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
  • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
  • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
  • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)

Key features and details

  • Rabbit polyclonal to KDM5C / Jarid1C / SMCX
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

You may also be interested in

Biochemical
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X-Gal, chromogenic beta-galactosidase substrate (ab144388)
Protein
Product image
Recombinant human KDM5C / Jarid1C / SMCX protein (ab196392)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

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: 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

    • WB: HEK-293T, HAP1 and Y79 cell lysates. ICC: HeLa cells.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Co-activators/co-repressors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Co-factors
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Jumonji containing proteins

Associated products

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • KO cell lines

    • Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (ab266251)
  • KO cell lysates

    • Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell lysate (ab257494)
  • Recombinant Protein

    • Recombinant human KDM5C / Jarid1C / SMCX protein (ab196392)

Applications

The Abpromise guarantee

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
WB (3)
1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 176 kDa).

Abcam recommends using milk as the blocking agent.

ICC/IF (1)
Use a concentration of 1 µg/ml.
Notes
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.
  • Target information above from: UniProt accession P41229 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8242 Human
    • Entrez Gene: 100037295 Pig
    • Omim: 314690 Human
    • SwissProt: P41229 Human
    • Unigene: 631768 Human
    • 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

    • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)

      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.

    • Immunocytochemistry - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Immunocytochemistry - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)

      ab34718 staining KDM5C / Jarid1C / SMCX in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab34718 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    • Immunocytochemistry/ Immunofluorescence - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Immunocytochemistry/ Immunofluorescence - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)

      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

    • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      All lanes : Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1/250 dilution

      Lane 1 : HEK-293T cell lysate
      Lane 2 : KDM5C knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 176 kDa
      Observed band size: 175 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab34718 observed at 175 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

      ab34718 was shown to react with KDM5C / Jarid1C / SMCX in wild-type HEK-293T cells in western blot with loss of signal observed in KDM5C knockout cell line ab266251 (KDM5C knockout cell lysate ab257494). Wild-type and KDM5C knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab34718 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 250 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      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.

    • Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
      Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1/250 dilution + HEK-293 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

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (7)

    Publishing research using ab34718? Please let us know so that we can cite the reference in this datasheet.

    ab34718 has been referenced in 7 publications.

    • Zhang B  et al. KDM5C Represses FASN-Mediated Lipid Metabolism to Exert Tumor Suppressor Activity in Intrahepatic Cholangiocarcinoma. Front Oncol 10:1025 (2020). PubMed: 32714863
    • Hong Z  et al. KDM5C is transcriptionally regulated by BRD4 and promotes castration-resistance prostate cancer cell proliferation by repressing PTEN. Biomed Pharmacother 114:108793 (2019). PubMed: 30921702
    • Wang J  et al. Asymmetric Expression of LincGET Biases Cell Fate in Two-Cell Mouse Embryos. Cell 175:1887-1901.e18 (2018). PubMed: 30550787
    • 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 . 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). PubMed: 21996408
    • Huang PH  et al. Histone deacetylase inhibitors stimulate histone H3 lysine 4 methylation in part via transcriptional repression of histone H3 lysine 4 demethylases. Mol Pharmacol 79:197-206 (2011). WB, ChIP ; Human . PubMed: 20959362
    • Kim JH & Youn HD C-terminal binding protein maintains mitochondrial activities. Cell Death Differ 16:584-92 (2009). ChIP ; Human . PubMed: 19136938

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-KDM5C / Jarid1C / SMCX antibody

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Embryonic Stem Cells)
    Gel Running Conditions
    Non-reduced Denaturing (8%)
    Loading amount
    50 µg
    Specification
    Embryonic Stem Cells
    Blocking step
    Odyssey PBS blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 30 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-KDM5C / Jarid1C / SMCX antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Read More

    DR. Kirk Mcmanus

    Verified customer

    Submitted Feb 27 2015

    Western blot abreview for Anti-KDM5C / Jarid1C / SMCX antibody

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 29 2013

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-KDM5C / Jarid1C / SMCX antibody

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

    Abcam user community

    Verified customer

    Submitted May 30 2012

    Western blot abreview for Anti-KDM5C / Jarid1C / SMCX antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 12 2008

    Question

    I am trying to see if this antibody could also react with the Y paralogue for this gene, Kdm5d. If you have any information on this please let me know.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 26 2012

    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

    Abcam Scientific Support

    Answered on Jun 26 2012

    Question

    I entered information for an abreview for this product last month. For some reason the review shows up as blank on the site even though I filled out the form, and I can't submit a new one because I already submitted the earlier one. I assume I can still use the code for a free antibody since a review was submitted, but I would like to submit one with the actual info and a picture if possible to put the information out there. Let me know if there's a way to do that please. Sorry about that, I'm not sure why that happened.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 25 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

    Abcam Scientific Support

    Answered on Apr 25 2012

    Question

    We have successfully used the antibody in mouse samples. I would be happy to send some pictures-will that allow us to then use the code? Also, we would like to validate the antibody, so I'm wondering if the antigen is available for purchase.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 24 2012

    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

    Abcam Scientific Support

    Answered on Feb 24 2012

    Question

    I am detecting a band at ˜80kDa instead of 176kDa. Is it possible to try another aliquot of antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 16 2012

    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 Scientific Support

    Answered on Feb 16 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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