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Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Histone H3.
(Peptide available as ab4557.)
Our Abpromise guarantee covers the use of ab28940 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K79) peptide (ab4557).|
|ChIP||Use 4-6 µg for 25 µg of chromatin.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab28940 specifically recognises a band at 15 kDa corresponding to methylated histone H3 (lane 1) that is blocked using mono- (ab4555 lane 2), di- (ab4556 lane 3) and tri-methylated (ab4557 lane 4) histone K79 H3 peptides. ab28940 is not blocked by di-methylated K79 H4 peptide (ab4560 lane 5), tri-methylated K4 H3 peptide (ab1342 lane 6) or unmodified H3 peptide (ab4558 lane 7).
This indicates that ab28940 specifically recognises all Histone H3 methylated K79 modifications.
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 ab28940 (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.
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