Overview

  • Product name
    Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8]
    See all KDM5A / Jarid1A / RBBP2 primary antibodies
  • Description
    Mouse monoclonal [18E8] to KDM5A / Jarid1A / RBBP2
  • Specificity
    Specific to human and mouse RBBP2. Can detect endogenous levels of RBBP2.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human KDM5A/ Jarid1A/ RBBP2 aa 1416-1434.
    Sequence:

    PRKQPRKSPLVPRSLEPPV


    Database link: P29375

  • Positive control
    • HeLa siRNA. HeLa RBP2 siRNA. Extracts from MCF7 cells, U2OS cells, NIH3T3 cells and J1 (mouse ES) cells.
  • General notes

    Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code.

Properties

Applications

Our Abpromise guarantee covers the use of ab78322 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
Flow Cyt Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
ICC/IF Use a concentration of 1 - 10 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 196 kDa.

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



  • Predicted band size : 196 kDa

    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 - ab78322 observed at 240 kDa. Red - loading control, ab176560, observed at 50 kDa.

    ab78322 was shown to specifically react with KDM5A when KDM5A knockout samples were used. Wild-type and KDM5A knockout samples were subjected to SDS-PAGE. Ab78322 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml 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.

     

     

  • ab78322 staining KDM5A / Jarid1A / RBBP2 [18E8] in HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4 % paraformaldehyde, permeabilized with Triton X-100 0.25% in PBS and blocked with 1.5% BSA for 30 minutes. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for overnight at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/1000 in PBS + 1% BSA) was used as the secondary antibody and incubated for 1 hour at room temperature. DAPI was used to stain the nuclear DNA.

  • All lanes : Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322) at 1 µg/ml

    Lane 1 : HeLa control siRNA
    Lane 2 : HeLa RBBP2 siRNA
    Lane 3 : Extracts from MCF7 cells
    Lane 4 : Extracts from U2OS cells
    Lane 5 : Extracts from NIH3T3 cells
    Lane 6 : Extracts from J1 (mouse ES) cells

    Developed using the ECL technique

    Predicted band size : 196 kDa
    Observed band size : 196 kDa
  • Overlay histogram showing HeLa cells stained with ab78322 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab78322, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Penterling C  et al. Depletion of Histone Demethylase Jarid1A Resulting in Histone Hyperacetylation and Radiation Sensitivity Does Not Affect DNA Double-Strand Break Repair. PLoS One 11:e0156599 (2016). WB, IF . Read more (PubMed: 27253695) »
  • Yamamoto S  et al. JARID1B is a luminal lineage-driving oncogene in breast cancer. Cancer Cell 25:762-77 (2014). WB . Read more (PubMed: 24937458) »

See all 5 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab78322.
Please use the links above to contact us or submit feedback about this product.

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

Sign up