GSM7157852: Zbtb46GFP_Flt3L+IL-6_2; Mus musculus; OTHER
The stimulated 5 x 105 cells were harvested and adsorbed to activated Concanavalin A (ConA) beads, permeabilized with 0.01% digitonin, and incubated with 0.5 mg C/EBPβ antibody or mouse IgG2a isotype control antibody at 4 °C on a nutator overnight. After antibody binding, the target-DNA complex is cleaved by the Protein A/Protein G-Micrococc al Nuclease (pAG-MNase) for 2 hours at 4 °C. 0.05 ng E. coli Spike-in DNA was added before releasing CUT&RUN fragments at 37°C for 10 min. The CUT&RUN enriched DNA is collected and purified from the supernatant. Libraries were prepared with a NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs) and cleaned-up with AMPureXP.