Anti mouse and rabbit igg hrp conjugated secondary antibodies

Whole-cell yeast extracts were prepared using the trichloroacetic acid method as previously described (Chen et al., 2012 (link)). The pelleted cells from 5 ml of culture were washed once with water and resuspended in 10% trichloroacetic acid. The cells were lysed by vortexing with glass beads, and the protein lysates were pelleted by centrifugation at 12,000 g for 15 min. The pellets were washed with ice-cold 80% acetone, and proteins were dissolved in ×2 SDS sample loading buffer by boiling for 5 min. The samples were centrifuged for 5 min at 12,000 g, and the supernatant was retained as protein extract. The samples were resolved on 8% SDS-PAGE gel and transferred onto a polyvinylidene difluoride membrane (Immobilon-P; Millipore) using a semi-dry method. Anti-Myc and anti-FLAG antibodies were purchased from MBL. GAPDH was purchased from GeneTex. Anti-mouse and rabbit IgG HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology. Blots were developed using the Western Blotting substrate (Bio-Rad).

Whole‐cell yeast extracts were prepared using a trichloroacetic acid (TCA) method, as previously described.^[36] Human cells were lysed in NETN buffer (10 × 10⁻³m Tris‐HCl, pH 8.0, 100 × 10⁻³m NaCl, 1 × 10⁻³m EDTA, and 0.5% NP‐40) with protease inhibitors (Roche) on ice for 30 min. Samples were resolved on an 8% or 12% SDS‐PAGE gel and transferred onto a PVDF membrane (Immobilon‐P; Millipore) using a semi‐dry method. Anti‐HA and anti‐FLAG antibodies were purchased from MBL and Sigma, respectively. Anti‐His and anti‐GST antibodies were obtained from Abclone. GAPDH was purchased from GeneTex. Mouse anti‐TAF1 (6B3), anti‐Myc (9E10) and anti‐p21 (sc‐6246) antibodies were bought from Santa‐Cruz. The antibodies against H4K5ac, H4K8ac, H4K12 or H4K16ac were purchased from Millipore. Anti‐mouse and rabbit IgG HRP‐conjugated secondary antibodies were purchased from Santa Cruz Biotechnology. Blots were developed using the Western Blotting substrate (Bio‐Rad).

Whole cell extracts were prepared using a trichloroacetic acid (TCA) method as previously described (11 (link)). Samples were resolved on an 8% or 12% SDS-PAGE gel and transferred onto a PVDF membrane (Immobilon-P; Millipore) using a semi-dry method. Anti-Myc and anti-HA antibody antibodies were purchased from MBL. Anti-FLAG antibody for Western blot was obtained from Sigma (F1804). Anti-mouse and rabbit IgG HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology. Blots were developed using the Western Blotting substrate (Bio-Rad).