These primary antibodies were used: mouse anti-BrdU antibody clones: B44 and 3D4 (BD Biosciences), Bu20a and MoBu-1 (Exbio Praha), BMC9318 (Roche) and chicken polyclonal anti-BrdU antibody (Abcam). For the concurrent localisation of DNA synthesis and cellular proteins, we used these antibodies: mouse anti-SC35 antibody (Abcam), rabbit anti-H1.2 antibody (Abcam), mouse anti-coilin antibody (Abcam), mouse anti-mitochondrial antibody (MTC02 antibody; Abcam), mouse anti-NTH1 antibody (Abcam), mouse anti-RPA32 antibody (Abcam), mouse anti-PRAF1 antibody (Abcam), rabbit anti-PCNA antibody (Abcam), and mouse anti-MCM7 antibody (Abcam).
The following secondary antibodies were used: DyLight 649 anti-mouse, Alexa Fluor® 488 anti-mouse, Alexa Fluor® 488 anti-rabbit and DyLight 649 anti-chicken antibodies (Jackson ImmunoResearch).
The primary antibodies were diluted either in 25 mM Tris-HCl, pH = 7.5, 150 mM NaCl or in 1× buffer for exonuclease III. The secondary antibodies were diluted in 25 mM Tris-HCl, pH = 7.5 and 150 mM NaCl. The cells were incubated with primary and secondary antibodies for 30 minutes at 37°C (if exonuclease III was used) or at room temperature (RT). After washing, the cells were mounted in the solution of 90% glycerol, 50 mM Tris-HCl, pH 8.0 and 2.5% 1,4-diazabicyclo[2.2.2]octane and evaluated.
The following secondary antibodies were used: DyLight 649 anti-mouse, Alexa Fluor® 488 anti-mouse, Alexa Fluor® 488 anti-rabbit and DyLight 649 anti-chicken antibodies (Jackson ImmunoResearch).
The primary antibodies were diluted either in 25 mM Tris-HCl, pH = 7.5, 150 mM NaCl or in 1× buffer for exonuclease III. The secondary antibodies were diluted in 25 mM Tris-HCl, pH = 7.5 and 150 mM NaCl. The cells were incubated with primary and secondary antibodies for 30 minutes at 37°C (if exonuclease III was used) or at room temperature (RT). After washing, the cells were mounted in the solution of 90% glycerol, 50 mM Tris-HCl, pH 8.0 and 2.5% 1,4-diazabicyclo[2.2.2]octane and evaluated.