Product nameAnti-HDAC2 antibody - ChIP Grade
See all HDAC2 primary antibodies
DescriptionRabbit polyclonal to HDAC2 - ChIP Grade
Tested applicationsSuitable for: WB, IP, IHC-P, Dot blot, ICC/IF, ChIPmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Chicken
- WB: nuclear extract of HeLa human epithelioid carcinoma cell line. Immunoprecipitation: whole lysate of NIH 3T cells. IHC-P: human lymph node sections.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Constituent: 1% BSA
Concentration information loading...
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.
ChIP Related Products
Our Abpromise guarantee covers the use of ab7029 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000 - 1/20000. Predicted molecular weight: 55.3 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-P||1/500. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
|Dot blot||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|ChIP||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.
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.
- Information by UniProt
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
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.
Immunoprecipitation analysis of HeLa whole cell lysates, ab7029 was used to precipitate HDAC3 from solution.
Lane 1 : Anti-HDAC2 antibody - ChIP Grade (ab7029) at 5 µl
Lane 2 : Anti-HDAC2 antibody - ChIP Grade (ab7029) at 10 µl
Lane 3 : Negative Control
All lanes : HeLa whole cell lysate.
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
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.
This product has been referenced in:
- Campbell AE et al. NuRD and CAF-1-mediated silencing of the D4Z4 array is modulated by DUX4-induced MBD3L proteins. Elife 7:N/A (2018). Read more (PubMed: 29533181) »
- Czimmerer Z et al. The Transcription Factor STAT6 Mediates Direct Repression of Inflammatory Enhancers and Limits Activation of Alternatively Polarized Macrophages. Immunity 48:75-90.e6 (2018). Read more (PubMed: 29343442) »