Anti-HDAC2抗体[3F3] - ChIP Grade (ab51832)

概述

性能

应用

Our Abpromise guarantee covers the use of ab51832 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 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IP Use at an assay dependent concentration.

Use 10ug for 10-50ug cell lysate.

ChIP Use at an assay dependent concentration.

Use 10ug for 25ug of DNA

ICC/IF 1/25 - 1/100.

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.
    Forms transcriptional repressor complexes by associating with MAD, SIN3, YY1 and N-COR. Interacts in the late S-phase of DNA-replication with DNMT1 in the other transcriptional repressor complex composed of DNMT1, DMAP1, PCNA, CAF1. Deacetylates TSHZ3 and regulates its transcriptional repressor activity.
  • 组织特异性Widely expressed; lower levels in brain and lung.
  • 序列相似性Belongs to the histone deacetylase family. HD type 1 subfamily.
  • 翻译后修饰S-nitrosylated by GAPDH. In neurons, S-Nitrosylation at Cys-262 and Cys-274 does not affect the enzyme activity but abolishes chromatin-binding, leading to increases acetylation of histones and activate genes that are associated with neuronal development. In embryonic cortical neurons, S-Nitrosylation regulates dendritic growth and branching.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • D10Wsu179e antibody
    • HD 2 antibody
    • HD2 antibody
    • HDAC 2 antibody
    • Hdac2 antibody
    • HDAC2_HUMAN antibody
    • Histone deacetylase 2 (HD2) antibody
    • Histone deacetylase 2 antibody
    • OTTHUMP00000017046 antibody
    • OTTHUMP00000227077 antibody
    • OTTHUMP00000227078 antibody
    • RPD3 antibody
    • transcriptional regulator homolog RPD3 antibody
    • YAF1 antibody
    • YY1 associated factor 1 antibody
    • YY1 transcription factor binding protein antibody
    • Yy1bp antibody
    see all

Anti-HDAC2 antibody [3F3] - ChIP Grade 图像

  • ab51832 stained MCF7 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51832 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-HDAC2 antibody [3F3] - ChIP Grade (ab51832) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 5 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
    Lane 6 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)


    Exposure time : 16 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab51832 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • Chromatin was prepared from the cell lysate of rat liver, cultured hepatoma cells according to the Abcam X-ChIP protocol.  Cells were fixed with formaldehyde for 10 min. The primary antibody was diluted 1/100 and incubated with the sample for 1.5 hours at 25°C.  Anti RNA polymerase II was used on the positive control, whilst normal mouse IgG was used in the negative control, to compare difference of HDAC2 binding on silencer DNA and promoter DNA of a gene. Data is normalized to 10% Input. The immunoprecipitated DNA was quantified by real time PCR.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab51832 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab51832, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-HDAC2 antibody [3F3] - ChIP Grade (ab51832)参考文献

This product has been referenced in:
  • Liu J  et al. MTA1 regulates higher-order chromatin structure and histone H1-chromatin interaction in-vivo. Mol Oncol 9:218-35 (2015). Read more (PubMed: 25205035) »
  • Rei D  et al. Basolateral amygdala bidirectionally modulates stress-induced hippocampal learning and memory deficits through a p25/Cdk5-dependent pathway. Proc Natl Acad Sci U S A 112:7291-6 (2015). WB ; Mouse . Read more (PubMed: 25995364) »

See all 8 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Raji, RcK8)
Gel Running Conditions Reduced Denaturing (10)
Loading amount 20 µg
Specification Raji, RcK8
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Xiao Qing Lu

Verified customer

提交于 Jan 25 2016

Thank you for contacting us.

The brownish or pinkish colour is very normal with antibodies if they are not purified. The ab51832 is also in tissue culture supernatant so the solution colour may be different due to presence of culture medium. <...

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Thank you for contacting us. I apologize for the inconvenience this may have caused.

I have had chat with goods-in department; unfortunately the in-stock vials are no longer available so it is not possible for us to manually check the vials. T...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Schwann cell)
Loading amount 200 µg
Specification Schwann cell
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°°C
Username

Mr. Tom Hu

Verified customer

提交于 Mar 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Mouse Tissue lysate - whole (Schwann cell)
Specification Schwann cell
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Paraformaldehyde
Detection step Semiquantitative PCR
Positive control Control
Negative control HDAC1/2 cko
Username

Mr. Tom Hu

Verified customer

提交于 Mar 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (lymphocyte)
Loading amount 500000 cells
Specification lymphocyte
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Jan 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Cultured hepatoma cell line)
Specification Cultured hepatoma cell line
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 27 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Mouse Cell lysate - whole cell (Liver, Hepatoma)
Specification Liver, Hepatoma
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step Real-time PCR
Positive control Anti RNA polymerase II
Negative control Normal mouse IgG
Username

Abcam user community

Verified customer

提交于 Sep 18 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Rat Cell lysate - whole cell (Liver, cultured hepatoma cells)
Specification Liver, cultured hepatoma cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step Real-time PCR
Positive control Anti RNA polymerase II
Negative control Normal mouse IgG
Username

Abcam user community

Verified customer

提交于 Jul 25 2008

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