Mouse anti xpress

After three rounds of MACS, the enriched SNU475 yeast display library was induced for FACS. From the first to the third round of FACS, 0.2 × 10 7 -0.8 × 10 7 yeast cells were sorted through FACSCalibur equipped with the sorting accessories (BD Bioscience, San Jose, CA, USA). In the fourth round of FACS, 3.1 × 10 7 yeast cells were run through FACS AriaIII (BD Bioscience).
The induced yeast cells in 100 µl of PBSF (0.8% NaCl, 0.02% KCl, 0.144% Na 2 HPO 4 , 0.024% KH 2 PO 4 , 0.1% BSA, and 0.0744% EDTA-2Na) were labeled with 1 µg of mouse anti-Xpress (Invitrogen) and 5 µg of polyclonal rabbit anti-actin (Invitrogen) antibodies. Two-color labeling was performed with 1 µg of FITC-conjugated donkey anti-mouse IgG (Jackson Labs, PA, USA) and 1 µg of PEconjugated goat anti-rabbit IgG (Santa Cruz Biotechnology, CA, USA). Samples thus prepared could be used for FACS and IF. For IF, an aliquot of the stained yeast cells was loaded onto a glass slide with a cover-glass. The sorted yeast cells were grown in SD-CAA medium at 30°C for 3 days.