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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

Reviews (1) Submit a question References (4)
Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Immunocytochemistry/ Immunofluorescence - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Immunoprecipitation - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Flow Cytometry (Intracellular) - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
  • Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18651] to KDM5A / Jarid1A / RBBP2
  • Suitable for: ICC/IF, IP, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651]
    See all KDM5A / Jarid1A / RBBP2 primary antibodies

  • Description

    Rabbit monoclonal [EPR18651] to KDM5A / Jarid1A / RBBP2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IP, WB, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, LCC, HEK-293 and NIH/3T3 whole cell lysates; mouse testis lysate. ICC/IF: HeLa cells. IP: HeLa whole cell lysate. Flow Cyt (intra): NIH/3T3 cell lysate

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab194286 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
1/2000.
IP
1/100.
WB
1/5000. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa).
Flow Cyt (Intra)
1/1500.
Notes
ICC/IF
1/2000.
IP
1/100.
WB
1/5000. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa).
Flow Cyt (Intra)
1/1500.

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'. May stimulate transcription mediated by nuclear receptors. May be involved in transcriptional regulation of Hox proteins during cell differentiation. May participate in transcriptional repression of cytokines such as CXCL12.

  • Sequence similarities

    Belongs to the JARID1 histone demethylase family.
    Contains 1 ARID domain.
    Contains 1 JmjC domain.
    Contains 1 JmjN domain.
    Contains 3 PHD-type zinc fingers.

  • Domain

    The GSGFP motif is required for the interaction with SUZ12.

  • Cellular localization

    Nucleus > nucleolus. Occupies promoters of genes involved in RNA metabolism and mitochondrial function.
  • Information by UniProt

  • Database links

    • Alternative names

      • Histone demethylase JARID1A antibody
      • JARID1A antibody
      • Jumonji/ARID domain containing protein 1A antibody
      • Jumonji/ARID domain-containing protein 1A antibody
      • Kdm5a antibody
      • KDM5A_HUMAN antibody
      • Lysine-specific demethylase 5A antibody
      • RBBP-2 antibody
      • RBBP2 antibody
      • RBP2 antibody
      • Retinoblastoma binding protein 2 antibody
      • Retinoblastoma-binding protein 2 antibody
      see all

    Images

    • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: KDM5A knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab194286 observed at 240 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab194286 was shown to specifically recognize KDM5A in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KDM5A knockout samples were examined. Wild-type and KDM5A knockout samples were subjected to SDS-PAGE. Ab194286 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/2500 dilution and 1/10000 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.

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Immunocytochemistry/ Immunofluorescence - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KDM5A / Jarid1A / RBBP2 with ab194286 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

      The negative controls are as follows:
      -ve control 1: ab194286 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
      -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

    • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      All lanes : Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286) at 1/5000 dilution

      Lane 1 : LLC (Mouse lung carcinoma) whole cell lysate
      Lane 2 : Mouse testis lysate
      Lane 3 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 192 kDa
      Observed band size: 192 kDa


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286) at 1/5000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 192 kDa
      Observed band size: 192 kDa


      Exposure time: 10 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286) at 1/1000 dilution + NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 192 kDa
      Observed band size: 192 kDa


      Exposure time: 10 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Immunoprecipitation - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

      KDM5A / Jarid1A / RBBP2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab194286 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab194286 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HeLa whole cell lysate 10ug (Input).

      Lane 2: ab194286 IP in HeLa whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194286 in HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 5 seconds.

    • Flow Cytometry (Intracellular) - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Flow Cytometry (Intracellular) - Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

      Intracellular Flow Cytometry analysis of NIH/3T3 (mouse embryo) cells labelling KDM5A / Jarid1A / RBBP2 (red) with purified ab194286 at dilution of 1/1500. The secondary antibody used was Alexa Fluorr® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody used was Rabbit Monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody. 

       

       

    • Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)
      Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286)

    References (4)

    Publishing research using ab194286? Please let us know so that we can cite the reference in this datasheet.

    ab194286 has been referenced in 4 publications.

    • Khaliq M  et al. Epigenetic modulation reveals differentiation state specificity of oncogene addiction. Nat Commun 12:1536 (2021). PubMed: 33750776
    • Zheng L  et al. ELK4 promotes the development of gastric cancer by inducing M2 polarization of macrophages through regulation of the KDM5A-PJA2-KSR1 axis. J Transl Med 19:342 (2021). PubMed: 34372882
    • Oser MG  et al. The KDM5A/RBP2 histone demethylase represses NOTCH signaling to sustain neuroendocrine differentiation and promote small cell lung cancer tumorigenesis. Genes Dev 33:1718-1738 (2019). PubMed: 31727771
    • McBrayer SK  et al. Autochthonous tumors driven by Rb1 loss have an ongoing requirement for the RBP2 histone demethylase. Proc Natl Acad Sci U S A N/A:N/A (2018). PubMed: 29610306

    Customer reviews and Q&As

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    ChIP abreview for Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651]

     Inconclusive
    Abreviews
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (K562)
    Specification
    K562
    Detection step
    Other
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Andrew Perez

    Verified customer

    Submitted Mar 24 2022

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