SpecificityAnti-Histone Deacetylase 2 specifically recognizes histone deacetylase 2 by immunoblotting and immunoprecipitation (55 kD).
The sequence recognized by the antibody is highly conserved in mouse and chicken.
Mouse, Human Predicted to work with:
Synthetic peptide corresponding to Human HDAC2 aa 471-488 conjugated to Keyhole Limpet Haemocyanin (KLH). The epitope recognized by the antibody is resistant to routine formalin-fixation and paraffin-embedding. Sequence:
Purification notesWhole antiserum is fractionated and further purified by anion-exchange chromatography to provide the IgG fraction of antiserum that is essentially free of other rabbit serum proteins.
1/500. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
Use at an assay dependent concentration. PubMed: 16713578
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. 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 specificityWidely expressed; lower levels in brain and lung.
Sequence similaritiesBelongs to the histone deacetylase family. HD type 1 subfamily.
Post-translational modificationsS-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.
Immunoprecipitation analysis of HeLa whole cell lysates, ab7029 was used to precipitate HDAC3 from solution.
Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody - ChIP Grade (ab7029)This image is courtesy of Michael Mancini, Baylor College of Medicine
HeLa cells were fixed with 4% formaldehyde in PEM buffer. The coverslip was incubated in blocking buffer of 5% powdered milk in TBS-T plus 0.02% sodium azide for 1 hour at room temperature. Blocking buffer was removed and primary antibody was added at a dilution of 1/600 and incubated overnight at 4 degrees celsius. The coverslips were then washed 4-5 times with blocking buffer for 5 minutes. Secondary antibody, goat anti-rabbit Alexa 594, was added at a dilution of 1/1000 and incubated at room temperature for one hour. From this point on coverslips were covered with foil to protect them from light. They were washed 5 times with TBS-T and then one time with PEM, for 5 minutes each wash. The coverslips were fixed 10-30 minutes in 4% formaldehyde in PEM buffer, then washed 3 times with PEM buffer for 5 minutes. 0.1M ammonium chloride in PEM buffer was added for 10 minutes to quench auto-florescence, and then slips were washed 2 times for 5 minutes in PEM followed by 3 washes for 5 minute
ChIP - HDAC2 antibody - ChIP Grade (ab7029)This image is courtesy of an anonymous Abreview
Chromatin was prepared from mouse skin epidermis whole tissue lysate. Cells were fixed with 1% fromaldehyde for 10 minutes. Wild type animal samples were incubated with rabbit IgG or ab7029 (diluted 1/800) for 12 hours at 4°C and the immunoprecipitated DNA was amplified using primers flanking the proximal promoter region and quantified by semiquantitative PCR. Rabbit IgG did not produce significant amplification.
HDAC2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HDAC2 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 ab7029. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: Band: 60ka: HDAC2; 55kDa; Heavy Chain.
References for Anti-HDAC2 antibody - ChIP Grade (ab7029)
This product has been referenced in:
Mould AW et al. Blimp1/Prdm1 Functions in Opposition to Irf1 to Maintain Neonatal Tolerance during Postnatal Intestinal Maturation. PLoS Genet11:e1005375 (2015).
Read more (PubMed: 26158850) »
Heckman LD et al. Rett-causing mutations reveal two domains critical for MeCP2 function and for toxicity in MECP2 duplication syndrome mice. Elife3:N/A (2014).
Read more (PubMed: 24970834) »