ChIP-Seq of AbFlex Histone H3.3 antibody (rAb) ChIP was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from brain tumor cell cells and 4 ug of AbFlex Histone H3.3 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15.1 million sequence tags were mapped to identify Histone H3.3 binding sites. The image shows binding across a region of chromosome 1.
AbFlex Histone H3.3 antibody (rAb) tested by Western Blot Lane 1: 250 ng of Recombinant Histone H3.1 (Cat. #31294); Lane 2: 250 ng of Recombinant Histone H3.3 (Cat. # 31295). Proteins were run on SDS-PAGE, transferred to nitrocellulose and stained with Ponceau S (lower panel) or probed with AbFlex Histone H3.3 antibody (upper panel). The AbFlex antibody only detects, and is specific for, Histone H3.3 as seen by a band at 17 kDa. For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.