Synthetic peptide corresponding to Human HDAC3 aa 411-428 conjugated to Keyhole Limpet Haemocyanin (KLH). The epitope recognized by the antibody is resistant to routine formalin-fixation and paraffin-embedding. Sequence:
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).
Use at an assay dependent concentration.
Use 2µl for 106 cells. PubMed: 18830415
Use at an assay dependent concentration.
Responsible 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. Probably participates in the regulation of transcription through its binding to the zinc-finger transcription factor YY1; increases YY1 repression activity. Required to repress transcription of the POU1F1 transcription factor. Acts as a molecular chaperone for shuttling phosphorylated NR2C1 to PML bodies for sumoylation.
Belongs to the histone deacetylase family. HD type 1 subfamily.
Western blot - Anti-HDAC3 antibody - ChIP Grade (ab7030)
All lanes : Anti-HDAC3 antibody - ChIP Grade (ab7030) at 1/5000 dilution
Lane 1 : HEK-293T whole cell lysate. Lane 2 : HeLa whole cell lysate. Lane 3 : K562 whole cell lysate. Lane 4 : COS-7 whole cell lysate. Lane 5 : CHO whole cell lysate. Lane 6 : NIH-3T3 whole cell lysate. Lane 7 : Neuro-2a whole cell lysate. Lane 8 : PC-12 whole cell lysate. Lane 9 : NRK whole cell lysate.
Secondary All lanes : Goat Anti-Rabbit IgG-Peroxidase and a chemiluminescent substrate.
Predicted band size: 49 kDa
ChIP - Anti-HDAC3 antibody - ChIP Grade (ab7030)This image is courtesy of Sylvain Daujat
Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab7030 to HDAC3 and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 2 µl of ab7030 and 2x10
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody - ChIP Grade (ab7030)This image is courtesy of an anonymous Abreview
Immunohistochemical analysis of paramaldehyde fixed human breast tumor using ab7030 at 1/500 dilution,followed by secondary antibody.
Immunocytochemical immunofluorescence analysis of HeLa cells labelling HDAC3 with ab7030. Cells were fixed and permeabilized with cold methanol followed by a cold methanol and acetone solution. Fixed cells were stained with ab7030 at a 1/200 concentration. The secondary used was a Goat Anti-Rabbit IgG, Cy3 conjugate.
Western blot - Anti-HDAC3 antibody - ChIP Grade (ab7030)This image is courtesy of an anonymous Abreview
All lanes : Anti-HDAC3 antibody - ChIP Grade (ab7030) at 1/10000 dilution
Lane 1 : Whole cell lysate from human HEK293 cell line Lane 2 : Whole cell lysate from human HEK293 cell line treated with HDAC3 gene silencing shRNA
Lysates/proteins at 20 µg per lane.
Secondary All lanes : HRP-conjugated goat anti-rabbit Ig at 1/5000 dilution
Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody - ChIP Grade (ab7030)This image is courtesy of Michael Mancini, Baylor College of Medicine
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 1/5000 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 minutes in
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