Recombinant Anti-HIRA/HIR antibody [EPR7416] (ab129169)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7416] to HIRA/HIR
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HIRA/HIR antibody [EPR7416]
See all HIRA/HIR primary antibodies -
Description
Rabbit monoclonal [EPR7416] to HIRA/HIR -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK293 and HeLa cell lysates ICC/IF: K562 and HeLa cells Flow Cyt (intra): HeLa and HEK293 cells IP: HEK293 cell lysates
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 1.35 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7416 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB | (1) |
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) |
1/10 - 1/100.
|
ICC/IF | (3) |
1/50 - 1/200.
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Notes |
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Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
ICC/IF
1/50 - 1/200. |
Target
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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
modificationsSumoylated.
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
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Database links
- Entrez Gene: 7290 Human
- Omim: 600237 Human
- SwissProt: P54198 Human
- Unigene: 474206 Human
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Alternative names
- DGCR1 antibody
- DiGeorge critical region gene 1 antibody
- HIR antibody
see all
Images
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All lanes : Anti-HIRA/HIR antibody [EPR7416] (ab129169) at 1/1000 dilution (purified)
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 minutesWe recommend to use 1%SDS Hot lysis prepare method.
We are unsure how to define the extra bands.
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Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HIRA/HIR with purified 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 Fluorr® 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).
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Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling HIRA/HIR with purified 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 ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (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.
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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 (+): ab129169 & HEK-293 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab129169 in HEK-293 whole cell lysate
For western blotting, 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.
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Overlay histogram showing HEK293 cells stained with unpurified 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.
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ab129169 (unpurified) at 1/50 dilution, staining HIRA/HIR in HeLa cells by Immunofluorescence.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab129169 has been referenced in 3 publications.
- Tachiwana H et al. Chromatin structure-dependent histone incorporation revealed by a genome-wide deposition assay. Elife 10:N/A (2021). PubMed: 33970102
- Gomes AP et al. Dynamic Incorporation of Histone H3 Variants into Chromatin Is Essential for Acquisition of Aggressive Traits and Metastatic Colonization. Cancer Cell 36:402-417.e13 (2019). PubMed: 31564638
- 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 . PubMed: 27939217