Anti-HDAC2 antibody [HDAC2-62] - ChIP Grade (ab12169)

Overview

  • Product name
    Anti-HDAC2 antibody [HDAC2-62] - ChIP Grade
    See all HDAC2 primary antibodies
  • Description
    Mouse monoclonal [HDAC2-62] to HDAC2 - ChIP Grade
  • Tested applications
    Suitable for: ChIP, ICC/IF, IHC-Fr, IHC-P, ELISA, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human
  • Immunogen

    Synthetic peptide:

    SGEKTDTKGTKSEQLSNP

    with N-terminally added Cysteine conjugated to KLH, corresponding to amino acids 471-488 of Human HDAC2.

  • Positive control
    • Whole cell extracts from NIH-3T3 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab12169 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
ChIP Use at an assay dependent concentration. PubMed: 24919190
ICC/IF 1/50.
IHC-Fr 1/1000. PubMed: 20407577
IHC-P Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 0.25 - 0.5 µg/ml. Detects a band of approximately 55 kDa.

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. 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.
  • Tissue specificity
    Widely expressed; lower levels in brain and lung.
  • Sequence similarities
    Belongs to the histone deacetylase family. HD type 1 subfamily.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • Chromatin was isolated and pooled from the two PF cortices of a single mouse. ChIP experiments were performed using a minimum of 3 mice per group. Chromatin was sheared in an ice bath by a 25 cycles of 30 sec on/off sonication using the “Bioruptor UCD-20” sonicator. Samples were kept on ice during 30 s between two pulses. An aliquot of precleared chromatin was collected as the input. The samples were incubated overnight at 4°C with 5 µl ab12169. The immunoprecipitated chromatin was analyzed by quantitative PCR with primers designed to amplify short regions of the promoters of genes of interest.

    ChIP assay on c-Fos gene promoter using ab12169 antibody. Results show a significant lower enrichment of HDAC 2 in PF cortex of (amyloid precursor protein) APP−/− mouse, Student’s t-test: **p<0.01. IgG was used as negative control. Equal amounts of ChIP and input chromatin were used for qRT-PCR analysis on the c-Fos gene promoter. Enrichment values were normalized to input values and represented the average of three or more mice per experiment. Results are expressed as mean ± SD.

  • ab12169 staining HDAC2 in a human glioblastoma cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 (0.1%) in PBS and blocked with 0.5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/50 in PBS + 0.5% BSA) for 16 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse monoclonal (1/400) was used as the secondary antibody. Nuclei were counterstained with Hoechst (blue).

    See Abreview

  • Western Blot using Rabbit anti-HDAC2

  • All lanes : Anti-HDAC2 antibody [HDAC2-62] - ChIP Grade (ab12169) at 0.5 µg/ml

    Lane 1 : Hek293T cell Lysate
    Lane 2 : HeLa cell Lysate
    Lane 3 : Jurkat cell Lysate
    Lane 4 : K562 cell Lysate
    Lane 5 : Neuro-2a cell Lysate
    Lane 6 : NIH-3T3 cell Lysate

    Secondary
    Goat Anti-Mouse IgG-Peroxidase
  • Anti-HDAC2 antibody [HDAC2-62] - ChIP Grade (ab12169) at 1/250 dilution + Human glioblastoma cell line at 120 µg

    Secondary
    HRP-conjugated Goat anti-Mouse at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 45 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 5 minutes

    This image is courtesy of an anonymous abreview.

    See Abreview

References

This product has been referenced in:
  • Rodnoi P  et al. Neuropeptide Y expression marks partially differentiated ß cells in mice and humans. JCI Insight 2:N/A (2017). ChIP ; Mouse . Read more (PubMed: 28614797) »
  • Yu T  et al. CBX7 is a glioma prognostic marker and induces G1/S arrest via the silencing of CCNE1. Oncotarget 8:26637-26647 (2017). WB . Read more (PubMed: 28460453) »

See all 33 Publications for this product

Customer reviews and Q&As

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

Abcam user community

Verified customer

Submitted Dec 11 2017

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

Abcam user community

Verified customer

Submitted May 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line)
Specification
human glioblastoma cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 11 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (lymphoma, multiple myeloma, prostate, ovarian)
Specification
lymphoma, multiple myeloma, prostate, ovarian
Fixative
Formaldehyde
Permeabilization
Yes - Target Retrieval Solution
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Username

Dr. Lena Marquard

Verified customer

Submitted Aug 01 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (U936 cell line (monocytic))
Specification
U936 cell line (monocytic)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Feb 20 2006

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

Sign up