Nucleosome fractions containing 200 ng DNA were applied for SDS-PAGE
with 4-20 % gradient SDS-PAGE gel (Bio-rad). Proteins were transferred to
the nitrocellulose membrane (GE) from the SDS-PAGE gel using TE42 Tank
Blotting Units (Hoefer) at 15 V, 4 °C for 4 h. As primally
antibodies, 1 μg/ml of Mouse monoclonal H3T3ph antibody 16B2 (Kelly et al., 2010 (link)) and 1 μg/ml
of mouse monoclonal PCNA antibody (Santa Cruz SC-56) were used. For H4 and
H1.8 detection, antibodies described above were used as primally antibodies.
As secondary antibodies, IRDye 680LT goat anti-rabbit IgG (Li-Cor 926-68021;
1:10,000) and IR Dye 800CW goat anti-mouse IgG (Li-Cor 926-32210; 1:15,000)
were used. The images were taken with Odyssey Infrared Imaging System
(Li-Cor).
with 4-20 % gradient SDS-PAGE gel (Bio-rad). Proteins were transferred to
the nitrocellulose membrane (GE) from the SDS-PAGE gel using TE42 Tank
Blotting Units (Hoefer) at 15 V, 4 °C for 4 h. As primally
antibodies, 1 μg/ml of Mouse monoclonal H3T3ph antibody 16B2 (Kelly et al., 2010 (link)) and 1 μg/ml
of mouse monoclonal PCNA antibody (Santa Cruz SC-56) were used. For H4 and
H1.8 detection, antibodies described above were used as primally antibodies.
As secondary antibodies, IRDye 680LT goat anti-rabbit IgG (Li-Cor 926-68021;
1:10,000) and IR Dye 800CW goat anti-mouse IgG (Li-Cor 926-32210; 1:15,000)
were used. The images were taken with Odyssey Infrared Imaging System
(Li-Cor).