概述

  • 产品名称Anti-HDAC1抗体
    参阅全部 HDAC1 一抗
  • 描述
    兔多克隆抗体to HDAC1
  • 经测试应用适用于: ICC/IF, WB, IP, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, African Green Monkey
    预测可用于: Cow
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human HDAC1.

    (Peptide available as ab20434.)

  • 阳性对照
    • This antibody gave a positive signal in Hela whole cell lysate and Human Breast Carcinoma.

性能

应用

Our Abpromise guarantee covers the use of ab19845 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 55 kDa).
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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
  • 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

Anti-HDAC1 antibody 图像

  • ICC/IF image of ab19845 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab19845, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    Panel A shows localisation of ab19845 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.



  • Predicted band size : 55 kDa

    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 - ab19845 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab19845 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. ab19845 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • The image shows staining of human tonsil tissue using ab19845. Staining was nuclear and was equally successful using Tris EDTA pH9 or Citrate pH6 for antigen retrieval. Staining was prevalent in almost all cellular compartments of the tonsil.
  • All lanes : Anti-HDAC1 antibody (ab19845) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa whole cell lysate with Human HDAC1 peptide (ab20434) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/5000 dilution

    Predicted band size : 55 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)
  • ab19845 at a 1/3000 dilution staining asynchronous HeLa cells by ICC/IF. The cells were paraformaldehyde fixed and immunofluorescently labelled with ab19845 for 30 minutes at room temperature. Bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody. Nuclei were visuallised using DAPI staining. The antibody was found to be highly enriched in the nucleus.

    This image is courtesy of an Abreview submitted by Kirk McManus.

    See Abreview

  • ICC/IF image of ab19845 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19845, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml, and in 100% methanol fixed (5 min) MCF7 and HepG2 cells at 1µg/ml
  • IHC image of HDAC1 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab19845, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
  • HDAC1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HDAC1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19845.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 60ka: HDAC1.

Anti-HDAC1 antibody (ab19845)参考文献

This product has been referenced in:
  • Lutz L  et al. Histone modifiers and marks define heterogeneous groups of colorectal carcinomas and affect responses to HDAC inhibitors in vitro. Am J Cancer Res 6:664-76 (2016). WB, IHC-P ; Human . Read more (PubMed: 27152243) »
  • Benard A  et al. Nuclear expression of histone deacetylases and their histone modifications predicts clinical outcome in colorectal cancer. Histopathology 66:270-82 (2015). IHC-P ; Human . Read more (PubMed: 25307864) »

See all 22 Publications for this product

Product Wall

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Sample African Green Monkey Cultured Cells (COS7)
Specification COS7
Permeabilization Yes - 0.5% TritonX100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jan 29 2014

Application Western blot
Sample Mouse Tissue lysate - other (Sciatic nerve and spinal cord)
Loading amount 10 µg
Specification Sciatic nerve and spinal cord
Gel Running Conditions Reduced Denaturing (8-12%)
Blocking step blocking buffer (infrared imaging system) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Dr. Gillian Hunter

Verified customer

提交于 Mar 26 2012

Thank you foryour reply. I am sorry that this antibody did not perform as stated on the datasheet. As requested, I have asked our Finance department to issue a credit note for you:

Credit note ID: 20243

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Read More
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (spinal cord)
Specification spinal cord
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Sep 06 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (various cell types - cell lines and human primary)
Loading amount 15 µg
Specification various cell types - cell lines and human primary
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Prof. Paul Townsend

Verified customer

提交于 Apr 21 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (HeLa cells)
Loading amount 10 µg
Specification HeLa cells
Gel Running Conditions Reduced Denaturing (7)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Mar 08 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (aortic smooth muscle cells)
Loading amount 5 µg
Specification aortic smooth muscle cells
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Feb 26 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (Endothelial cells)
Loading amount 4.5 µg
Specification Endothelial cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 07 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

提交于 Jun 05 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"