Anti sir2 yn 19

Proteins from mid-log phase cultures were isolated using trichloroacetic acid as described previously[18 (link)]. The samples were resolved by electrophoresis on a 10% SDS-polyacrylamide gel. The following primary antibodies were used for immunoblotting at a 1:1000 dilution: Anti-Pgk1 (ab113678), anti-Rad53 (ab104232), and anti γ-H2A (ab15083) were purchased from Abcam; anti-HA (#2578) was purchased from Cell Signaling; anti-Myc (sc-40), anti-Hda1 (sc-6657), anti-Rpd3 (yC-19), and anti-Sir2 (yN-19) were purchased from Santa Cruz Biotechnology; anti-GFP (G1544) was purchased from Sigma-Aldrich; anti-mouse (A9044), anti-rabbit (A0545), and anti-goat (A5420) HRP-linked secondary antibodies were purchased from Sigma-Aldrich and used at a 1:100000 dilution. Images were acquired with a Wealtec KETA-CL imaging system.

The preparation of yeast protein extract from TCA-treated cells was described previously [57 (link)]. The samples were separated by electrophoresis on a 10% SDS-polyacrylamide gel. The following primary antibodies were used for immunoblotting at a 1:1000 dilution: anti-Pgk1 (ab113678), anti-Rad53 (ab104232) and anti Y-H2A (ab15083) were obtained from Abcam; anti-Rpd3 (yC-19), anti-Hda1 (yC-20) and anti-Sir2 (yN-19) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and anti-HA (H9658) and anti-Myc (M4439) were obtained from Sigma-Aldrich. Anti-mouse (A9044), anti-rabbit (A0545) and anti-goat (A5420) HRP-linked secondary antibodies were obtained from Sigma-Aldrich (St. Louis, MO, USA) and used at a 1:100,000 dilution. Images were acquired with a Wealtec KETA-CL imaging system.