| Description |
293T cells were subjected to nuclear and cytoplasmic separation. The nucleus was then lysed and the resulting supernatant was collected. Dynabead MyOne Streptavidin C1 beads were mixed with the supernatant for incubation at 4 °C for 2 hours to eliminate background noise and recover the supernatant. Meanwhile, freshly washed C1 beads were mixed with peptides with different numbers of methyl groups and incubated at 4 °C for 4 hours. The beads were subsequently washed, mixed with the nucleoplasm supernatant and incubated overnight at 4 °C on a rotator. A portion of beads were eluted with SDS-PAGE loading buffer and subjected to silver staining for checking the eluted proteins. The rest of the beads were eluted with Urea-Elution buffer (10mM Tris-HCl, 6M Urea, 2M Thiourea, pH 8.0.). The eluted protein samples were loaded onto the stacking gel and resolved for 5cm. The gel lane that hold loaded proteins was then cut and submitted to the facility. |