使用敲除细胞株进行验证重组RabMAb

重组Anti-HDAC9抗体[EPR5223] (ab109446)

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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, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Rat, Human

概述

  • 产品名称

    Anti-HDAC9抗体[EPR5223]
    参阅全部 HDAC9 一抗

  • 描述

    兔单克隆抗体[EPR5223] to HDAC9

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt, WB, IHC-P, ICC/IFmore details
    不适用于: IP

  • 种属反应性

    与反应: Rat, Human

  • 免疫原

    Synthetic peptide within Human HDAC9 aa 1-100 (N terminal). The exact sequence is proprietary.

  • 阳性对照

    • IHC-P: Human colon Tissue ICC/IF: HeLa cells WB: HepG2, Raji, HAP1, cell lysates Flow Cyt: K-562 cells.

  • 常规说明

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt Use at an assay dependent concentration.
WB 1/10000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
  • 应用说明
    Is unsuitable for IP.

    • 靶标

    • 功能

      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.

    • 组织特异性

      Broadly expressed, with highest levels in brain, heart, muscle and testis. Isoform 3 is present in human bladder carcinoma cells (at protein level).

    • 疾病相关

      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.

    • 序列相似性

      Belongs to the histone deacetylase family. HD type 2 subfamily.

    • 翻译后修饰

      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.

    • 细胞定位

      Nucleus.
    • Information by UniProt

    • 数据库链接

      • 别名

        • 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

      图片

      • 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.


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

        ab109446 at 1/250 dilution staining HDAC9 in Human colon by Immunohistochemistry, Paraffin-embedded tissue.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Immunocytochemistry/ Immunofluorescence - Anti-HDAC9 antibody [EPR5223] (ab109446)
        Immunocytochemistry/ Immunofluorescence - Anti-HDAC9 antibody [EPR5223] (ab109446)
        ab109446 at 1/100 dilution staining HDAC9 in HeLa cells by Immunofluorescence.
      • Flow Cytometry - Anti-HDAC9 antibody [EPR5223] (ab109446)
        Flow Cytometry - Anti-HDAC9 antibody [EPR5223] (ab109446)

        Flow cytometry of K-562 Human chronic myelogenous leukemia lymphoblast cells staining HDAC9 with ab109446 (red) at 1:250. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used at 1:2000 as a secondary. Rabbit monoclonal IgG (ab172730)  (Black) was used as an isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) used as an unlabelled control. Fixed with 4% paraformaldehyde. Permeabilised with 90% methanol.

      • Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
        Western blot - Anti-HDAC9 antibody [EPR5223] (ab109446)
        All lanes : Anti-HDAC9 antibody [EPR5223] (ab109446) at 1/10000 dilution

        Lane 1 : K562 cell lysate
        Lane 2 : HepG2 cell lysate
        Lane 3 : Raji cell lysate

        Lysates/proteins at 10 µg per lane.

        Predicted band size: 111 kDa
        Observed band size: 160 kDa why is the actual band size different from the predicted?

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

      文献 (11)

      发表研究结果有使用 ab109446？请让我们知道，以便我们可以引用本数据表中的参考文章。

      ab109446 被引用在 11 文献中.

      • Li H  et al. High HDAC9 is associated with poor prognosis and promotes malignant progression in pancreatic ductal adenocarcinoma. Mol Med Rep 21:822-832 (2020). PubMed: 31974610
      • Fu Y  et al. Decreased expression and hypomethylation of HDAC9 lead to poor prognosis and inhibit immune cell infiltration in clear cell renal cell carcinoma. Urol Oncol N/A:N/A (2020). PubMed: 32402768
      • 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
      • Luo L  et al. HDAC4 Controls Muscle Homeostasis through Deacetylation of Myosin Heavy Chain, PGC-1a, and Hsc70. Cell Rep 29:749-763.e12 (2019). PubMed: 31618641
      • Li A  et al. Histone deacetylase 6 regulates the immunosuppressive properties of cancer-associated fibroblasts in breast cancer through the STAT3-COX2-dependent pathway. Oncogene N/A:N/A (2018). PubMed: 29980788
      View all Publications for this product

      客户评价及客户问答

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      1-2 of 2 Abreviews or Q&A

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

       Excellent
      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

      Mrs. Alessandra Granata

      Verified customer

      提交于 Feb 06 2020

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

       Excellent
      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

      Courtney Fisher

      Verified customer

      提交于 Apr 19 2017

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