Overview

  • Product name
    Anti-HIRA antibody [EPR7416]
    See all HIRA primary antibodies
  • Description
    Rabbit monoclonal [EPR7416] to HIRA
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human HIRA aa 950 to the C-terminus. The exact sequence is proprietary.
    Database link: P54198

  • Positive control
    • 293T, HeLa and K562 cell lysates, HeLa cells
  • General notes

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab129169 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 1/1000 - 1/10000. Predicted molecular weight: 112 kDa.

We recommend to use 1%SDS Hot lysis prepare method. For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

IP 1/10 - 1/100.
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/50 - 1/100.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Cooperates with ASF1A to promote replication-independent chromatin assembly. Required for the periodic repression of histone gene transcription during the cell cycle. Required for the formation of senescence-associated heterochromatin foci (SAHF) and efficient senescence-associated cell cycle exit.
    • Tissue specificity
      Expressed at high levels in kidney, pancreas and skeletal muscle and at lower levels in brain, heart, liver, lung, and placenta.
    • Sequence similarities
      Belongs to the WD repeat HIR1 family.
      Contains 8 WD repeats.
    • Developmental stage
      Expressed during embryogenesis.
    • Post-translational
      modifications
      Sumoylated.
      Phosphorylated by CDK2/CCNA1 and CDK2/CCNE1 on Thr-555 in vitro. Also phosphorylated on Thr-555 and Ser-687 in vivo.
    • Cellular localization
      Nucleus. Nucleus > PML body. Primarily, though not exclusively, localized to the nucleus. Localizes to PML bodies immediately prior to onset of senescence.
    • Information by UniProt
    • Database links
    • Alternative names
      • DGCR1 antibody
      • DiGeorge critical region gene 1 antibody
      • HIR antibody
      • HIR histone cell cycle regulation defective homolog A antibody
      • HIRA antibody
      • HIRA protein antibody
      • HIRA_HUMAN antibody
      • Histone cell cycle regulator antibody
      • Protein HIRA antibody
      • TUP 1 antibody
      • TUP1 like enhancer of split protein 1 antibody
      • TUP1-like enhancer of split protein 1 antibody
      • TUPLE1 antibody
      see all

    Images

    • All lanes : Anti-HIRA antibody [EPR7416] (ab129169) at 1/1000 dilution

      Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
      Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method with 5% NFDM/TBST

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 112 kDa
      Observed band size: 112 kDa


      Exposure time: 3 minutes


      We recommend to use 1%SDS Hot lysis prepare method.

      We are unsure how to define the extra bands.

    • Overlay histogram showing HEK293 cells stained with ab129169 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab129169, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • ab129169, at 1/50 dilution, staining HIRA in HeLa cells by Immunofluorescence.
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:
    • Zhu Z  et al. PHB Associates with the HIRA Complex to Control an Epigenetic-Metabolic Circuit in Human ESCs. Cell Stem Cell 20:274-289.e7 (2017). WB ; Human . Read more (PubMed: 27939217) »
    See 1 Publication for this product

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton X-100
    Specification
    HeLa
    Fixative
    Paraformaldehyde

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Aug 10 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HAP1)
    Permeabilization
    Yes - 0.5% Triton 5 minutes
    Specification
    HAP1
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Feb 13 2018

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

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