使用敲除细胞株进行验证重组RabMAb

重组Anti-HDAC1抗体[EPR460(2)] (ab109411)

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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 [EPR460(2)] to HDAC1
  • Suitable for: IHC-P, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-HDAC1抗体[EPR460(2)]
    参阅全部 HDAC1 一抗

  • 描述

    兔单克隆抗体[EPR460(2)] to HDAC1

  • 宿主

    Rabbit

  • 经测试应用

    适用于: IHC-P, WB, Flow Cytmore details
    不适用于: IP

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human HDAC1 aa 400-500 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab174565)

  • 阳性对照

    • WB: Jurkat, HeLa, K562, and MCF7 cell lysates. IHC-P: Human colon and tonsil tissues.

  • 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 ab109411 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
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/1000 - 1/10000. Predicted molecular weight: 55 kDa.
Flow Cyt 1/50.
  • 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. Histone deacetylases act via the formation of large multiprotein complexes. Deacetylates SP proteins, SP1 and SP3, and regulates their function. Component of the BRG1-RB1-HDAC1 complex, which negatively regulates the CREST-mediated transcription in resting neurons. Upon calcium stimulation, HDAC1 is released from the complex and CREBBP is recruited, which facilitates transcriptional activation. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. Deacetylates 'Lys-310' in RELA and thereby inhibits the transcriptional activity of NF-kappa-B.

    • 组织特异性

      Ubiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.

    • 序列相似性

      Belongs to the histone deacetylase family. HD type 1 subfamily.

    • 翻译后修饰

      Sumoylated on Lys-444 and Lys-476; which promotes enzymatic activity. Desumoylated by SENP1.
      Phosphorylation on Ser-421 and Ser-423 promotes enzymatic activity and interactions with NuRD and SIN3 complexes.
      Ubiquitinated by CHFR, leading to its degradation by the proteasome.

    • 细胞定位

      Nucleus.
    • Information by UniProt

    • 数据库链接

      see all

    • 别名

      • DKFZp686H12203 antibody
      • GON 10 antibody
      • HD1 antibody
      • HDAC 1 antibody
      • HDAC1 antibody
      • HDAC1_HUMAN antibody
      • Histone deacetylase 1 antibody
      • Reduced potassium dependency yeast homolog like 1 antibody
      • RPD3 antibody
      • RPD3L1 antibody
      see all

    图片

    • Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: HDAC1 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Human breast carcinoma lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109411 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109411 was shown to recognize HDAC1 when HDAC1 knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC1 knockout samples were subjected to SDS-PAGE. ab109411 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] (ab109411)

      Immunohistochemical analysis of HDAC1 in paraffin-embedded Human colon tissue using ab109411 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Flow Cytometry - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Flow Cytometry - Anti-HDAC1 antibody [EPR460(2)] (ab109411)

      Overlay histogram showing HeLa cells stained with ab109411 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab109441, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22°C.

      Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.1ug/1x106 cells) used under the same conditions.

      Acquisition of >5,000 events were collected using an 50 mW Blue laser (488nm) with 530/30 bandpass filter.

    • Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: HDAC1 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Human breast carcinoma lysate (20 µg) or K562 lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109411 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

      This western blot image is a comparison between ab109411 and a competitor's top cited rabbit polyclonal antibody.

    • Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      All lanes : Anti-HDAC1 antibody [EPR460(2)] (ab109411) at 1/1000 dilution

      Lane 1 : Mouse brain lysate, 20ug
      Lane 2 : Mouse heart lysate, 20ug
      Lane 3 : Mouse Kidney lysate, 20ug
      Lane 4 : Mouse spleen lysate,20ug

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated(Pierce). at 1/1000 dilution

      Predicted band size: 55 kDa
      Observed band size: 62 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and Diluting buffer: 5% NFDM /TBST

    • Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Western blot - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      All lanes : Anti-HDAC1 antibody [EPR460(2)] (ab109411) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : K562 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 55 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] (ab109411)

      Immunohistochemical analysis of HDAC1 in paraffin-embedded Human tonsil tissue using ab109411 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-HDAC1 antibody [EPR460(2)] (ab109411)
      Anti-HDAC1 antibody [EPR460(2)] (ab109411)

    文献 (16)

    发表研究结果有使用 ab109411？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab109411 被引用在 16 文献中.

    • Xiong W  et al. MiR-761 inhibits colorectal cancer cell proliferation and invasion through targeting HDAC1. Pharmazie 74:111-114 (2019). PubMed: 30782261
    • Bartosz P  et al. Benzophenone-3 passes through the blood-brain barrier, increases the level of extracellular glutamate and induces apoptotic processes in the hippocampus and frontal cortex of rats. Toxicol Sci N/A:N/A (2019). PubMed: 31368502
    • Loktev SS & Ogneva IV DNA Methylation of Mouse Testes, Cardiac and Lung Tissue During Long-Term Microgravity Simulation. Sci Rep 9:7974 (2019). PubMed: 31138883
    • King HW  et al. Polycomb repressive complex 1 shapes the nucleosome landscape but not accessibility at target genes. Genome Res 28:1494-1507 (2018). PubMed: 30154222
    • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
    View all Publications for this product

    客户评价及客户问答

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    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-HDAC1 antibody [EPR460(2)]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Brain)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    12 µg
    Treatment
    Aliquoted and heated from 34C to 69C to determine melting temperature of HDAC1. Lysate centrifuged at 18,000 x g to remove aggregates after melting. Supernatent collected, 3X SDS PAGE loading dye added, and heated sampled were heated to 95C for 10 min. Note, input sample was not heated before centrifugation, but was heated to 95C after loading dye added.
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 17 2015

    Western blot abreview for Anti-HDAC1 antibody [EPR460(2)]

     Inconclusive
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% Tris-bis glycine gel)
    Sample
    Mouse Tissue lysate - other (mouse brain tissue)
    Specification
    mouse brain tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Arjang Salehi

    Verified customer

    提交于 Mar 26 2015

    Western blot abreview for Anti-HDAC1 antibody [EPR460(2)]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - nuclear (B cells)
    Specification
    B cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 16 2015

    Western blot abreview for Anti-HDAC1 antibody [EPR460(2)]

     Average
    Abreviews
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - whole cell (heart)
    Specification
    heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 13 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-HDAC1 antibody [EPR460(2)]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 28°C
    Sample
    Mouse Cell (Neural cell)
    Specification
    Neural cell
    Permeabilization
    Yes - 0.3% TritonX-100 in PBS for 30 minutes
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Oct 13 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-HDAC1 antibody [EPR460(2)]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    提交于 May 31 2012

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