The H3K4me3 antibody is a rabbit polyclonal that was raised against a modified branched peptide to specifically recognize histone H3 tri-methylated on lysine 4 (H3K4me3). The antibody detects a single protein of approximately 14 kDa by western blot using acid extracted histones. The antibody has been tested in chromatin immunoprecipitation (ChIP) experiments against a strain lacking all H3K4 methyltransferase activity and determined to be specific.
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 the antibody to beads.
For optimal ChIP results, we recommend using H3K4me3 supercoated ChIP-Right beads which were derived by selection of a recombinant antibody library generated from splenocytes of immunized animals.