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  1. Link

    hdac9-antibody-epr5223-ab109446.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Acetylation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HDAC9 antibody [EPR5223] (ab109446)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (16)

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Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
  • Flow Cytometry (Intracellular) - Anti-HDAC9 antibody [EPR5223] (ab109446)
  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC9 antibody [EPR5223] (ab109446)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC9 antibody [EPR5223] (ab109446)
  • Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
  • Immunoprecipitation - Anti-HDAC9 antibody [EPR5223] (ab109446)
  • 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: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP PE

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Overview

  • Product name

    Anti-HDAC9 antibody [EPR5223]
    See all HDAC9 primary antibodies
  • Description

    Rabbit monoclonal [EPR5223] to HDAC9
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the IHC-P results. We do not guarantee WB for mouse and rat.

  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Rat and mouse cerebrum tissue. WB: HepG2, Raji and HAP1 whole cell lysate. IP: K-562 whole cell lysate.
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5223
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • HDACs
    • Class II / Hda1 Class
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-HDAC9 antibody [EPR5223] (ab206713)
    • Alexa Fluor® 647 Anti-HDAC9 antibody [EPR5223] (ab207050)
    • Anti-HDAC9 antibody [EPR5223] - BSA and Azide free (ab239979)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Hep G2 nuclear extract lysate (ab14660)
    • K-562 nuclear extract lysate (ab14851)
    • Raji whole cell lysate (ab30124)
  • Recombinant Protein

    • Recombinant human HDAC9 protein (ab80350)

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
Flow Cyt (Intra)
1/100.
IP
1/30.
WB
1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa).

The mouse and rat recommendation is based on the IHC-P results. We do not guarantee WB for mouse and rat.

IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF (1)
1/100 - 1/250.
Notes
Flow Cyt (Intra)
1/100.
IP
1/30.
WB
1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa).

The mouse and rat recommendation is based on the IHC-P results. We do not guarantee WB for mouse and rat.

IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.

Target

  • 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
    modifications

    Phosphorylated 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.
  • Target information above from: UniProt accession Q9UKV0 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 9734 Human
    • Entrez Gene: 79221 Mouse
    • Entrez Gene: 687001 Rat
    • Omim: 606543 Human
    • SwissProt: Q9UKV0 Human
    • SwissProt: Q99N13 Mouse
    • Unigene: 196054 Human
    • Unigene: 310551 Mouse
    • Unigene: 483009 Mouse
    see all
  • Alternative names

    • HD 7 antibody
    • HD 7B antibody
    • HD 9 antibody
    • HD7 antibody
    • HD7B antibody
    • HD9 antibody
    • HDAC 7 antibody
    • HDAC 7B antibody
    • HDAC 9 antibody
    • HDAC 9B antibody
    • HDAC 9FL antibody
    • HDAC antibody
    • HDAC7 antibody
    • HDAC7B antibody
    • HDAC9 antibody
    • HDAC9_HUMAN antibody
    • HDAC9B antibody
    • HDAC9FL antibody
    • HDRP antibody
    • Histone deacetylase 4/5 related protein antibody
    • Histone deacetylase 7 antibody
    • Histone deacetylase 7B antibody
    • Histone deacetylase 9 antibody
    • Histone deacetylase 9A antibody
    • Histone deacetylase related protein antibody
    • Histone deacetylase-related protein antibody
    • KIAA0744 antibody
    • MEF2 interacting transcription repressor MITR antibody
    • MEF2 interacting transcription repressor protein antibody
    • MEF2-interacting transcription repressor MITR antibody
    • MITR antibody
    see all

Images

  • Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
    Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
    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 (Intracellular) - Anti-HDAC9 antibody [EPR5223] (ab109446)
    Flow Cytometry (Intracellular) - Anti-HDAC9 antibody [EPR5223] (ab109446)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC9 antibody [EPR5223] (ab109446)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC9 antibody [EPR5223] (ab109446)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC9 antibody [EPR5223] (ab109446)
    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.

  • Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
    Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)

    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.

  • Immunoprecipitation - Anti-HDAC9 antibody [EPR5223] (ab109446)
    Immunoprecipitation - Anti-HDAC9 antibody [EPR5223] (ab109446)

    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.

  • Anti-HDAC9 antibody [EPR5223] (ab109446)
    Anti-HDAC9 antibody [EPR5223] (ab109446)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (16)

Publishing research using ab109446? Please let us know so that we can cite the reference in this datasheet.

ab109446 has been referenced in 16 publications.

  • 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
  • Xu L  et al. MicroRNA-936 inhibits the malignant phenotype of retinoblastoma by directly targeting HDAC9 and deactivating the PI3K/AKT pathway. Oncol Rep 43:635-645 (2020). PubMed: 31922233
  • Zaidi SAH  et al. Histone Deacetylases Regulation by d-Opioids in Human Optic Nerve Head Astrocytes. Invest Ophthalmol Vis Sci 61:17 (2020). PubMed: 32915982
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-2 of 2 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-HDAC9 antibody [EPR5223]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hiPSC-SMC (smooth muscle cells))
Permeabilization
Yes - 0.05% Triton-X 100
Specification
hiPSC-SMC (smooth muscle cells)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MRS. Alessandra Granata

Verified customer

Submitted Feb 06 2020

Immunohistochemistry (Frozen sections) abreview for Anti-HDAC9 antibody [EPR5223]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Basilar Artery)
Permeabilization
Yes - 0.1% Triton-X PBS
Specification
Basilar Artery
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Courtney Fisher

Verified customer

Submitted Apr 19 2017

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