Recombinant Anti-HDAC9 antibody [EPR5223] (ab109446)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5223] to HDAC9
- Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HDAC9 antibody [EPR5223]
See all HDAC9 primary antibodies -
Description
Rabbit monoclonal [EPR5223] to HDAC9 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, 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: HepG2, Raji, K-562, and HAP1 whole cell lysate. IHC-P: Human cerebrum tissue. ICC/IF: K-562 cells. Flow Cyt (Intra): K-562 cells. IP: K-562 whole cell lysate.
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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.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5223 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109446 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.
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IP |
1/30.
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WB |
1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/100. |
IP
1/30. |
WB
1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
Target
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Function
Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Represses MEF2-dependent transcription.
Isoform 3 lacks active site residues and therefore is catalytically inactive. Represses MEF2-dependent transcription by recruiting HDAC1 and/or HDAC3. Seems to inhibit skeletal myogenesis and to be involved in heart development. Protects neurons from apoptosis, both by inhibiting JUN phosphorylation by MAPK10 and by repressing JUN transcription via HDAC1 recruitment to JUN promoter. -
Tissue specificity
Broadly expressed, with highest levels in brain, heart, muscle and testis. Isoform 3 is present in human bladder carcinoma cells (at protein level). -
Involvement in disease
Note=A chromosomal aberration involving HDAC9 is found in a family with Peters anomaly. Translocation t(1;7)(q41;p21) with TGFB2 resulting in lack of HDAC9 protein. -
Sequence similarities
Belongs to the histone deacetylase family. HD type 2 subfamily. -
Post-translational
modificationsPhosphorylated on Ser-220 and Ser-450; which promotes 14-3-3-binding, impairs interaction with MEF2, and antagonizes antimyogenic activity. Phosphorylated on Ser-240; which impairs nuclear accumulation (By similarity). Isoform 7 is phosphorylated on Tyr-1010. Phosphorylated by the PKC kinases PKN1 and PKN2, impairing nuclear import.
Sumoylated. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 9734 Human
- Omim: 606543 Human
- SwissProt: Q9UKV0 Human
- Unigene: 196054 Human
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Alternative names
- HD 7 antibody
- HD 7B antibody
- HD 9 antibody
see all
Images
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All lanes : Anti-HDAC9 antibody [EPR5223] (ab109446) at 1/1000 dilution (Purified)
Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 111 kDa -
Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling HDAC9 with Purified ab109446 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). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling HDAC9 with Purified ab109446 at 1:100 dilution (10 µg/ml). Cells were fixed in 100% Methanol 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC9 antibody [EPR5223] (ab109446)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling HDAC9 with Purified ab109446 at 1:1000 dilution (1.10 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: HDAC9 (KO) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)
Lane 4: Raji whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109446 observed at 140 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109446 was shown to specifically recognize HDAC9 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when HDAC9 knockout samples were usedexamined. Wild-type and HDAC9 knockout samples were subjected to SDS-PAGE. Ab109446 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10,000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging. -
HDAC9 was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab109446 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 260035 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: K-562 whole cell lysate 10 μg
Lane 2: K-562 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab109446 in K-562 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 111 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (19)
ab109446 has been referenced in 19 publications.
- Lu Z et al. Promotion of microRNA-146a by histone deacetylase 4 silencing contributes to radiosensitization of esophageal carcinoma. J Transl Med 20:101 (2022). PubMed: 35193602
- Ye J et al. HDAC7 Activates IKK/NF-κB Signaling to Regulate Astrocyte-Mediated Inflammation. Mol Neurobiol 59:6141-6157 (2022). PubMed: 35871708
- Zhu Y & Hao F Knockdown of long non-coding RNA HCP5 suppresses the malignant behavior of retinoblastoma by sponging miR-3619-5p to target HDAC9. Int J Mol Med 47:N/A (2021). PubMed: 33693951
- Wan H et al. circ_0003204 Regulates Cell Growth, Oxidative Stress, and Inflammation in ox-LDL-Induced Vascular Endothelial Cells via Regulating miR-942-5p/HDAC9 Axis. Front Cardiovasc Med 8:646832 (2021). PubMed: 33869307
- Liu F et al. HDAC9 exacerbates myocardial infarction via inactivating Nrf2 pathways. J Pharm Pharmacol N/A:N/A (2021). PubMed: 33963859