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

Overview

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

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

  • Positive control
    • Ab46985 gave a positive signal in WB in the following whole cell lysates: HeLa; Jurkat; K562. This antibody gave a positive result in IHC in the following FFPE tissue: Human heart muscle.

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 Use 1µg for 106 cells.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 55 kDa).
IP Use at an assay dependent concentration.
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 Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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 antibody
    • GON 10 antibody
    • HD 1 antibody
    • HD1 antibody
    • HDAC 1 antibody
    • HDAC1 antibody
    • hdac1: histone deacetylase 1 antibody
    • HDAC1_HUMAN antibody
    • Histone deacetylase 1 (HD1) antibody
    • Histone deacetylase 1 antibody
    • Reduced potassium dependency yeast homolog like 1 antibody
    • RPD3 (reduced potassium dependency yeast homolog) like 1 antibody
    • RPD3 (reduced potassium dependency) antibody
    • RPD3 antibody
    • RPD3L1 antibody
    see all

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

  • All lanes : Anti-HDAC1 [10E2] antibody - ChIP Grade (ab46985) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate

    Lysates/proteins at 10 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 : 60 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes
  • IHC image of HDAC1 staining in Human heartmuscle 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 ab46985, 5µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • 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, 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. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • 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

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

This product has been referenced in:
  • Martin N  et al. Interplay between Homeobox proteins and Polycomb repressive complexes in p16INK4a regulation. EMBO J 32:982-95 (2013). Read more (PubMed: 23455154) »
  • Sharma D  et al. Dexamethasone induces a putative repressor complex and chromatin modifications in the CRH promoter. Mol Endocrinol 27:1142-52 (2013). Read more (PubMed: 23671328) »

See all 9 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Sample Rat Tissue sections (Brain)
Specification Brain
Permeabilization Yes - Citrate Buffer pH 6.0
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 20 2013

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Sample Mouse Tissue sections (mouse thyroid)
Specification mouse thyroid
Permeabilization No
Fixative Paraformaldehyde
Username

Mr. Sanjay Gawade

Verified customer

Submitted Jun 18 2013

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 go...

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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 ...

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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

Mr. Tom Hu

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

Mr. Tom Hu

Verified customer

Submitted Apr 03 2012

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