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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, acetylated at K27.
(Peptide available as ab24404.)
Our Abpromise guarantee covers the use of ab4729 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||1/500. Can be used with paraformaldehyde- or methanol- fixed cells.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (acetyl K27) peptide (ab24404).|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.|
|CHIPseq||Use at an assay dependent concentration.|
|ChIP/Chip||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
|PepArr||Use a concentration of 0.2 - 0.02 µg/ml.|
All batches of ab4729 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - acetyl K27 peptide (ab24404), indicating that this antibody specifically recognises the Histone H3 - acetyl K27 modification.
ab24404 - Histone H3 - acetyl K27
ab15591 - Histone H3 - acetyl K14
ab24003 - Histone H3 - acetyl K18
ab17163 - Histone H3 unmodified
ab48359 - Histone H3 - acetyl K23
ab41409 - Histone H3 - acetyl K36
ab15662 - Histone H4 - acetyl K12
ab16635 - Histone H3 acetyl K9
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 ab4729 (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.
Primary antibody: ab4729 (H3 acetyl K27)
ab4729 strongly stained histones of mouse ES cells. However, fluroescence was greatly diminished following pre-blocking using a H3 acetyl K9 peptide. This suggests the antibody cross-reacts with the K9 and K27 residues.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"