Mouse monoclonal anti myc sc 40

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.

Yeast cells were lysed with glass beads in 20% trichloroacetic acid. The pellet was washed and proteins were extracted with Laemmli buffer. Protein samples were denatured in boiling water for 5 min and separated by sodium dodecyl sulfate polyacrylamide gel . The following antibodies were used for immunoblotting after membrane transfer: mouse monoclonal anti-HA (H3663, Sigma), mouse monoclonal anti-Myc (sc-40, Santa Cruz Biotechnology), mouse monoclonal anti-Flag (66008-3, Proteintech), mouse monoclonal anti-ubiquitin (P4G7, Santa Cruz Biotechnology), rabbit monoclonal anti-ubiquitin (linkage-specific K48) (ab140601, Abcam), rabbit monoclonal anti-Myc (2278, Cell Signaling Technology), rabbit polyclonal anti-Flag (PM020, MBL), rabbit polyclonal anti-SUMO (gift from Wei Li, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China), and mouse monoclonal anti-PGK1 (ab113687, Abcam). Membranes were imaged with an Amersham Imager 680. Signal was quantified using Quantity One.