Pulldown of Dnmt1 using the Dnmt1-Trap: Immunoprecipitations (IP) of Dnmt1 from a protein extract of HEK 293T cells. Size of DNMT1: 186 kDA. Primary antibody: anti-DNMT1 1:500. Secondary antibody: anti-rat 1:2000. Input (I), non-bound (FT) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining and Western Blotting. Unlike conventional antibodies, the Dnmt1-Trap is free from heavy and/or light chain contaminants and therefore yields cleaner results.
Immunofluorescence: HeLa cells 3.7% PFA fixed. Primary antibody: 1/100 Dnmt1 antibody 2E8, incubation for 1 h at RT: Secondary antibody (top): 1/400 anti-rat-Alexa488, incubation for 1 h at RT. Secondary antibody (bottom): 1/400 anti-rat. Alexa568, incubation for 1 h at RT. Characteristic punctate staining of Dnmt1 at replication foci in the nucleus of cells in S phase.