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Synthetic peptide corresponding to Histone H3 aa 7-20 (N terminal). aa 7 to 20 and Ac-Lys9.
Our Abpromise guarantee covers the use of ab12179 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration.
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ChIP||Use 5-10 µg for 25 µg of chromatin.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 17 kDa.|
|ELISA||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
IHC image of ab12179 staining Histone H3 (acetyl K9) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab12179, 10µg/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. No primary antibody was used in the negative control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
SK-N-SH cells fixed in 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab12179 (1/1000 dilution). ab12179 staining is localized to the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.
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, 5µg of ab12179 (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.
ab12179 at a 1/1000 dilution staining Stylonychia lemnae (single cell organism, transcriptionally active macronucleus) by ICC/IF. The cells were paraformaldehyde fixed and incubated with the antibody for 12 hours. An Alexa Fluor ® 488 conjugated goat anti-mouse IgG antibody was used as the secondary.
In the image Histone H3 (acetyl K9) staining is red and is found in macronuclei only. Micronuclei remain unstained and are shown in blue (nucleic acid counterstain). Alpha tubulin is also stained (green).
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