GSM9453007: Jurkat cells, Empty, H3K27ac; Homo sapiens; OTHER
CUT&Tag assays were performed using the CUT&Tag Assay Kit (Cell Signaling Technology, #77552) according to the manufacturer's protocol with minor modifications. Briefly, 1×10^5 cells were fixed with 0.1% formaldehyde for 2 min at room temperature and quenched with 1.25 M glycine for 5 min. Cells were pelleted, washed once with Complete Wash Buffer containing spermidine and protease inhibitor cocktail, and resuspended in the same buffer. The cell suspension was incubated with Concanavalin A magnetic beads. Following bead binding, permeabilization, antibody incubation, tagmentation, and DNA purification were performed according to the manufacturer's protocol. Primary antibodies against H3K27ac and IRF4 were used. Libraries were prepared using the CUT&Tag Assay Kit (Cell Signaling Technology, #77552) according to the manufacturer's instructions.