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Synthetic peptide within Human Histone H3 aa 50 to the C-terminus (mono methyl K79) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
(Peptide available as
Our Abpromise guarantee covers the use of ab2886 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use 2µg for 106 cells.|
|ICC||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|Immunodiffusion||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 18787701|
|WB||1/1 - 1/500. Detects a band of approximately 17 kDa.|
|PepArr||Use a concentration of 0.2 - 0.02 µg/ml.|
All batches of ab2886 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 - mono methyl K79 peptide (ab4555), indicating that this antibody specifically recognises the Histone H3 - mono methyl K79 modification.
ab4558 - Histone H3 - unmodified
ab1771 - Histone H3 - di methyl K9
ab4555 - Histone H3 - mono methyl K79
ab4556 - Histone H3 - di methyl K79
ab4557 - Histone H3 - tri methyl K79
ab4560 - Histone H4 - di methyl K79
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab2886 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR. Primers and probes are located in the first kb of the transcribed region.
Chromatin was prepared from whole cell lysate of normal rat liver and liver cancer cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. 5 µg of the primary antibody was used in 1/100 dilution and it was incubated with the sample for 16 hours at 4°C in a commercially available ChIP dilution buffer. The immunoprecipitated DNA was quantified by real time PCR. ChIP results show that the Histone H3 (mono methyl K79) and GAPDH genes are expressed in higher levels in liver cancer cells than in normal liver cells.
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