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Rabbit Recombinant Monoclonal HIRA/HIR antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

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Images

Immunoprecipitation - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (AB232477), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (AB232477), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (AB232477), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (AB232477), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (AB232477), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)
Human
Tested
Expected
Tested
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

2 products for Alternative Version

    2 products for Alternative Product

      Target data

      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.

      Alternative names

      Protein HIRA, TUP1-like enhancer of split protein 1, TUPLE1, HIR, DGCR1, HIRA

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      Rabbit Recombinant Monoclonal HIRA/HIR antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

      Alternative names

      Protein HIRA, TUP1-like enhancer of split protein 1, TUPLE1, HIR, DGCR1, HIRA

      Key facts

      Isotype

      IgG

      Form

      Liquid

      Clonality

      Monoclonal

      Immunogen
      • The exact immunogen used to generate this antibody is proprietary information.
      Carrier free

      Yes

      Clone number

      EPR7416

      Purification technique

      Affinity purification Protein A

      Concentration
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      Storage

      Shipped at conditions

      Blue Ice

      Appropriate long-term storage conditions

      +4°C

      Storage information

      Do Not Freeze

      Notes

      ab232477 is the carrier-free version of Anti-HIRA/HIR antibody [EPR7416] ab129169.

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

      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 batch production

      For more information, read more on recombinant antibodies.

      Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

      This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

      Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

      This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

      Supplementary info

      Activity summary

      HIRA also known as HIR or HIR/HIRA is a histone chaperone protein that plays an essential role in chromatin remodeling and nucleosome assembly. This protein has a mass of approximately 121 kDa. It is expressed in various tissues with notable presence in the liver heart and skeletal muscle. The HIRA protein interacts with other proteins to regulate the deposition of histone H3.3 which is important for processes like DNA replication and repair.

      Biological function summary

      HIRA participates in the formation of a complex known as the HIRA-ASF1A-CABIN1 protein complex. This complex operates in chromatin assembly and modulates the incorporation of histone variant H3.3 affecting gene silencing and transcription regulation. Interacting with histone proteins HIRA serves an important function in genome stability and epigenetic mark establishment.

      Pathways

      HIRA influences the histone replacement pathway significantly impacting gene expression and chromatin organization. HIRA works closely with ASF1A and CABIN1 proteins within this pathway. Additionally in the senescence-associated secretory phenotype pathway HIRA contributes to cellular aging processes by regulating transcriptional outcomes. The modulation of these pathways by HIRA exemplifies its key role in maintaining cellular functions.

      Associated diseases and disorders

      Defects in HIRA associate with hypertrophic cardiomyopathy and some cancer types such as breast cancer. HIRA alterations can disrupt chromatin dynamics potentially linking it with poor prognostic factors in these diseases. In particular the HIRA gene can interact with other genes involved in cardiomyopathies like MYH7 affecting heart muscle function. Through its role in chromatin remodeling HIRA connects to various pathologies where gene regulation and DNA damage responses are compromised.

      Product promise

      We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

      In the unlikely event of one of our products not working as expected, you are covered by our product promise.

      Full details and terms and conditions can be found here:
      Terms & Conditions.

      6 product images

      • Immunoprecipitation - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477), expandable thumbnail

        Immunoprecipitation - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477)

        Anti-HIRA/HIR antibody [EPR7416] ab129169 (Purified) at 1/50 dilution (2ug) immunoprecipitating HIRA/HIR in HEK-293 whole cell lysate.
        Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10ug
        Lane 2 (+): Anti-HIRA/HIR antibody [EPR7416] ab129169 & HEK-293 whole cell lysate
        Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HIRA/HIR antibody [EPR7416] ab129169 in HEK-293 whole cell lysate
        For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/1000 dilution.
        Blocking and diluting buffer: 5% NFDM/TBST.

        This antibody works in the lysates prepared with 1%SDS Hot lysis method in WB. No target band in input lane is due to the lysates prepared with RIPA method.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIRA/HIR antibody [EPR7416] ab129169)

        All lanes: Immunoprecipitation - Anti-HIRA/HIR antibody [EPR7416] (Anti-HIRA/HIR antibody [EPR7416] ab129169)

        Predicted band size: 112 kDa

      • Flow Cytometry (Intracellular) - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477), expandable thumbnail

        Flow Cytometry (Intracellular) - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477)

        Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HIRA/HIR with purified Anti-HIRA/HIR antibody [EPR7416] ab129169 at 1/100 dilution (10μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIRA/HIR antibody [EPR7416] ab129169).

      • Immunocytochemistry/ Immunofluorescence - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477), expandable thumbnail

        Immunocytochemistry/ Immunofluorescence - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477)

        Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling HIRA/HIR with purified Anti-HIRA/HIR antibody [EPR7416] ab129169 at 1/200 dilution (5 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIRA/HIR antibody [EPR7416] ab129169).

      • Flow Cytometry (Intracellular) - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477), expandable thumbnail

        Flow Cytometry (Intracellular) - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477)

        Overlay histogram showing HEK293 cells stained with unpurifiedab129169 (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 (Anti-HIRA/HIR antibody [EPR7416] ab129169, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIRA/HIR antibody [EPR7416] ab129169).

      • Immunocytochemistry/ Immunofluorescence - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477), expandable thumbnail

        Immunocytochemistry/ Immunofluorescence - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477)

        Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling HIRA/HIR using Anti-HIRA/HIR antibody [EPR7416] ab129169 (unpurified), at 1/50 dilution.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIRA/HIR antibody [EPR7416] ab129169).

      • OI-RD Scanning - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477), expandable thumbnail

        OI-RD Scanning - Anti-HIRA/HIR antibody [EPR7416] - BSA and Azide free (ab232477)

        We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
        Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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      Product protocols

      For this product, it's our understanding that no specific protocols are required. You can:

      Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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