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Synthetic peptide within Human Histone H3 aa 50 to the C-terminus (tri methyl K79) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
(Peptide available as
Our Abpromise guarantee covers the use of ab2621 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|ChIP||Use 2-4 µg for 6 µg of chromatin.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K79) peptide (ab4557).
Abcam recommends using BSA as the blocking agent.
|ChIP/Chip||Use at an assay dependent concentration.|
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 ab2621 (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.
Immunocytochemistry/ Immunofluorescence analysis of HEK293 cells labeling Histone H3 (tri methyl K79) with ab2621 at 1/1000 dilution. Cells were fixed with paraformaldehyde and permeabilized with Triton-X. The cells were blocked with 5% BSA for 1 hour at 21°C, followed by incubation with Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) in 3% BSA in TBST for 16 hours at 4°C. A goat anti-rabbit Alexa Fluro 594 secondary antibody was used at 1/1000 dilution.
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. The immunoprecipitated DNA was quantified by real time PCR.