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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3.
(Peptide available as ab1773.) Clones were positively screened by ELISA against the immunising peptide conjugated to BSA. Clones were negatively screened against both the equivalent unmodified peptide conjugated to BSA and also a tri methyl K27 peptide conjugated to BSA.
Our Abpromise guarantee covers the use of ab6001 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 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K9) peptide (ab1773).|
|ICC/IF||Use a concentration of 5 µg/ml.|
ICC/IF image of ab6001 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Triton for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab6001 at 5µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-mouse DyLight® 488 (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All batches of ab6001 are tested in Peptide Array against peptides to different Histone H3 modifications at 20µg/ml. 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 - tri methyl K9 peptide (ab1773), indicating that this antibody specifically recognises the Histone H3 - tri methyl K9 modification.