Mouse monoclonal anti ha h9658

Yeast cells were lysed in 20% trichloroacetic acid using glass beads. The resultant pellets were extracted with a Laemmli buffer and denatured in boiling water for 5 min. Proteins were separated on a 10% SDS–polyacrylamamide gel electrophoresis gel. The membrane transfer was performed at a current of 200 mA on ice for 1.5 hours. The following antibodies were used for immunoblotting: mouse monoclonal anti-HA (H9658, Sigma-Aldrich), mouse monoclonal anti-Myc (sc-40, Santa Cruz Biotechnology), and mouse monoclonal anti-phosphoglycerate kinase 1 (PGK1; ab113687, Abcam). Membranes were imaged with a Bio-Rad Imager. Quantification of protein bands was performed using ImageJ software.