AbFlex Cas9 antibody (rAb) tested by Western blot. 50 ng recombinant Cas9 protein was probed with 0.5 ug/ml AbFlex Cas9 antibody (Lane 1). HEK293T cells were transfected with a plasmid containing a mammalian expression construct for dCas9 (S. pyogenes). Chromatin was prepared 48 hours post-transfection, boiled and 15 ul (~200,000 cell equivalents) was run on a SDS-PAGE gel and probed with 2 ug/ml AbFlex Cas9 antibody (Lane 2). A molecular weight marker was run in Lane 3.
AbFlex Cas9 antibody (rAb) tested by bead-based specificity analysis. Luminex bead-based specificity analysis was used to confirm the specificity of AbFlex Cas9 antibody (rAb) antibody for Cas9. Various proteins were conjugated to MagPlex Luminex beads and incubated with various amounts of AbFlex Cas9 antibody (rAb). Protein-bound antibody was detected with anti-mouse IgG-Phycoerythrin and read in a Luminex instrument. Luminex is a registered trademark of Luminex Corporation.
AbFlex Cas9 recombinant antibody (rAb) tested by ChIP-Seq Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of Jurkat cell chromatin and 4 ug of AbFlex Cas9 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 8 million sequence tags were mapped to identify binding sites on chromosome 4.
