The H3K27me3 antibody is a rabbit polyclonal that was raised against a branched peptide and recognizes histone H3 tri-methylated on lysine 27 (H3K27me3). The antibody detects a single protein of approximately 14 kDa by western blot using acid extracted histones. This antibody has been throughly tested in chromatin immunoprecipitation (ChIP) experiments.
ChIP Recommendations: Use 2.5-5.0 ul of crude sera with 20-30 ul of Apheon Protein A beads (or 40-60 ul of competitor beads) per 100 ug of sheared chromatin lysate. We recommend titrating the antibody, beads, and lysate to determine ideal concentrations. We also recommend pre-binding of the antibody to beads.
For optimal ChIP results, we recommend using H3K27me3 supercoated ChIP-Right beads which were derived by selection of a recombiant antibody library generated from splenocytes of immunized animals.