Anti-HDAC1 antibody (ab19845)
- Product nameAnti-HDAC1 antibodySee all HDAC1 primary antibodies ...
- DescriptionRabbit polyclonal to HDAC1
- Tested applicationsICC/IF, WB, IP, IHC-P more details
- Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Cow
Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human HDAC1.
(Peptide available as ab20434.)
- Positive control
- This antibody gave a positive signal in Hela whole cell lysate and Human Breast Carcinoma.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
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.
|ICC/IF||ICC/IF: Use a concentration of 1 µg/ml.|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 55 kDa).Can be blocked with Human HDAC1 peptide (ab20434).|
|IP||IP: Use at an assay dependent concentration.|
|IHC-P||IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
- 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.
modificationsSumoylated 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.
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Anti-HDAC1 antibody images
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.
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (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?)
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.
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.
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.
References for Anti-HDAC1 antibody (ab19845)
This product has been referenced in:
- Takase K et al. Monoaminergic and neuropeptidergic neurons have distinct expression profiles of histone deacetylases. PLoS One 8:e58473 (2013). ICC/IF ; Mouse . Read more (PubMed: 23469282) »
- Wells CE et al. Inhibition of histone deacetylase 3 causes replication stress in cutaneous T cell lymphoma. PLoS One 8:e68915 (2013). WB ; Mouse . Read more (PubMed: 23894374) »