Figure 1. Schematic overview of TAF-ChIP approach.

(1) Formaldehyde fixed cells were directly sorted into radio immunoprecipitation (RIPA) buffer (see the Materials and Methods section for details). (2) The cells were briefly sonicated at low intensity to break open the nuclei. (3) Antibodies were coupled to magnetic beads in the presence of blocking reagents. (4) Antibody-coupled beads were added to the cell lysate and incubated overnight at 4°C. (5) The tagmentation reaction was performed after initial washes with low salt IP buffer and homemade tagmentation buffer. (6) The tagmentation reaction and the background regions (not anchored by antibody interaction) were washed away with subsequent high-stringency washes. (7) The proteinase K was heat-inactivated and the material was PCR-amplified without purification.