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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, acetylated at K9.
Our Abpromise guarantee covers the use of ab10812 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ChIP||Use 2-4 µg for 25 µg of chromatin.|
|IP||Use at 20 µg/mg of lysate.|
|WB||1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (acetyl K9) peptide (ab16635).|
|CHIPseq||Use at an assay dependent concentration. PubMed: 22196736|
|ICC/IF||Use a concentration of 1 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration.|
ab10812 staining histone H3 (acetyl K9) in A549 cells treated with scriptaid (ab120883), by ICC/IF. Increase in histone H3 (acetyl K9) expression correlates with increased concentration of scriptaid, as described in literature.
The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab120883 (scriptaid) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab10812 (0.1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A anti-rabbit DyLight 488 secondary antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
ab10812 staining Histone H3 (acetyl K9) in Mouse thyroid tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/200 in 10% serum in PBS) for 1 hour at 24°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab10812 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
SK-N-SH cells fixed with 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab10812 (1/1000). ab10812 clearly stained the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.
ab10812 staining Histone H3 (acetyl K9) in the root tips of Arabidopsis thaliana by Immunocytochemistry/ Immunofluorescence. Cells were fixed in 2% paraformaldehyde for 30 minutes at room temperature. Blocking and permeabilization was carried with 4% BSA solution containing 0,5% Triton X-100 in PBS at room temperature for 45 minutes. Slides were washed in PBS and incubated with primary antibody at a 1/200 dilution in 1% PBS for 1 hour at 37°C. Slides were washed in PBS and incubated with the secondary antibody ab6639 (Goat anti-rabbit Cy3 ® (H&L)) at a 1/500 dilution in 1% BSA in PBS for 1 hour at 37°C. Slides were counterstained with DAPI (2µg/mL) for 10 minutes at room temperature.Left image: DAPI stained interphase nucleus with prominent chromocentersMiddle image: Distribution of Histone H3 (acetyl K9) in the nucleus (arrows indicate absence of signal from chromocenters)Right image: Merged image
Image courtesy of an anonymous Abreview.
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