Figure 2

Co-immunoprecipitation of the N-terminal domain of Dnmt1. COS-7 cells were transfected with Flag-tagged C-terminus (lanes 1, 4), Flag-tagged C-terminus + Flu-tagged N-terminus (lanes 2, 5) or with Flu-tagged N-terminus (lanes 3, 6). The cells were lysed in EBC buffer and the supernatants immunoprecipitated with α-Flag AB. The complexes were bound to agarose beads, washed and separated by electrophoresis. After transfer to PVDF membranes, the Western blots were probed with α-Flu AB, stripped and reprobed with α-Flag AB. Lanes 1–3 show a ~2% aliquot of the initial extract used in the immunoprecipitation (IP, lanes 4–6). The recovery rate of the immunoprecipitation was about 10 %.