Product nameAnti-HIRA/HIR antibody [EPR7416]
See all HIRA/HIR primary antibodies
DescriptionRabbit monoclonal [EPR7416] to HIRA/HIR
Tested applicationsSuitable for: WB, IP, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Human
within Human HIRA/HIR aa 950 to the C-terminus. The exact sequence is proprietary.
Database link: P54198
- WB: HEK293 and HeLa cell lysates ICC/IF: K562 and HeLa cells FC: HeLa and HEK293 cells IP: HEK293 cell lysates
This product was previously labelled as HIRA
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.
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.
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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 - 1/200.|
FunctionCooperates 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 specificityExpressed at high levels in kidney, pancreas and skeletal muscle and at lower levels in brain, heart, liver, lung, and placenta.
Sequence similaritiesBelongs to the WD repeat HIR1 family.
Contains 8 WD repeats.
Developmental stageExpressed during embryogenesis.
Phosphorylated by CDK2/CCNA1 and CDK2/CCNE1 on Thr-555 in vitro. Also phosphorylated on Thr-555 and Ser-687 in vivo.
Cellular localizationNucleus. Nucleus > PML body. Primarily, though not exclusively, localized to the nucleus. Localizes to PML bodies immediately prior to onset of senescence.
- Information by UniProt
- DGCR1 antibody
- DiGeorge critical region gene 1 antibody
- HIR antibody
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.
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.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HIRA/HIR with purified ab129169 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (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).
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.
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.
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.
ab129169 (unpurified) at 1/50 dilution, staining HIRA/HIR in HeLa cells by Immunofluorescence.
Equilibrium disassociation constant (KD)
Learn more about KD
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