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Anti-HDAC1 antibody [10E2] - ChIP Grade (ab46985)

Overview

  • Product nameAnti-HDAC1 antibody [10E2] - ChIP GradeSee all HDAC1 primary antibodies ...
  • Description
    Mouse monoclonal [10E2] to HDAC1 - ChIP Grade
  • SpecificityThis antibody recognizes HDAC1.
  • Tested applicationsFlow Cyt, WB, ICC/IF, IP, ChIP, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide: EEKPEAKGVKEEVKLA, corresponding to C terminal amino acids 467-482 of Human and mouse HDAC1

  • Positive controlHeLa cells. This antibody gave a positive result in IHC in the following FFPE tissue: Human heartmuscle.

Properties

Applications

Our Abpromise guarantee covers the use of ab46985 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Flow Cyt Flow Cyt: 1/100.
WB WB: 1/500 - 1/2000. Detects a band of approximately 57 kDa (predicted molecular weight: 55 kDa).
ICC/IF ICC/IF: 1/50 - 1/200.
IP IP: Use at an assay dependent dilution.
ChIP ChIP: 1/10 - 1/50. This antibody was validated for ChIP using 10 ul of antibody added to chromatin prepared from formaldehyde fixed U2OS cells. The p21 gene promoter was interrogated by PCR. The promoter of the HDAC1 gene was interrogated for the presence of HDAC1 from chromatin harvested from fixed Swiss 3T3 cells in separate experiements.
IHC-P IHC-P: Use a concentration of 5 µg/ml.

Target

  • FunctionResponsible 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.
  • Tissue specificityUbiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.
  • Sequence similaritiesBelongs to the histone deacetylase family. HD type 1 subfamily.
  • Post-translational
    modifications
    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.
  • Cellular localizationNucleus.
  • Target information above from: UniProt accession Q13547 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
      DKFZp686H12203 antibodyGON 10 antibodyHD 1 antibody
      HD1 antibodyHDAC 1 antibodyHDAC1 antibodyhdac1: histone deacetylase 1 antibodyHDAC1_HUMAN antibodyHistone deacetylase 1 (HD1) antibodyHistone deacetylase 1 antibodyReduced potassium dependency yeast homolog like 1 antibodyRPD3 (reduced potassium dependency yeast homolog) like 1 antibodyRPD3 (reduced potassium dependency) antibodyRPD3 antibodyRPD3L1 antibody
    see all

Anti-HDAC1 antibody [10E2] - ChIP Grade images

  • Anti-HDAC1 antibody [10E2] - ChIP Grade (ab46985) at 1/1000 dilution + Nuclear extract of HeLa cells.

    Predicted band size : 55 kDa
    Observed band size : 57 kDa (why is the actual band size different from the predicted?)
  • ChIP analysis using ab46985 binding HDAC1 in Schwann cells. Cells were fixed with paraformaldehyde for 10 mins. Samples were incubated with primary antibody for 12 hours at 4°C at an antibody concentration of 0.1µg/µl of chromatin. Protein binding was detected using semi-quantitative PCR at both a positive and negative control regions.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab46985 (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 (ab46985, 1/100 dilution) 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.

References for Anti-HDAC1 antibody [10E2] - ChIP Grade (ab46985)

This product has been referenced in:
  • Bantscheff M  et al. Chemoproteomics profiling of HDAC inhibitors reveals selective targeting of HDAC complexes. Nat Biotechnol : (2011). Read more (PubMed: 21258344) »
  • Opsahl JA  et al. Increased interaction between DJ-1 and the Mi-2/ nucleosome remodelling and deacetylase complex during cellular stress. Proteomics 10:1494-504 (2010). ICC/IF, WB ; Human . Read more (PubMed: 20127688) »

See all 6 Publications for this product

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Thank you for your enquiry regarding ab46985.

Some information is provided on the legend of the Flow image on the on-line product datasheet.

Blocking serum should always be from the same species as the secondary antibody - we use goat ...

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The concentration of this antibody has not been determined as it is derived from a tissue culture supernatant.

Thank you for your inquiry.

This antibody is a tissue culture supernatant, meaning that is an unpurified form. It is common for these products to be colored and/or cloudy. The antibody should work just fine. If you have any difficulty, please let...

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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 10 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Verified customer

Submitted Apr 18 2012

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
Negative control NA
Username

Verified customer

Submitted Apr 03 2012

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