pCT-NT-F2A

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	To induce cell surface protein expression, yeast cells containing pCTCON, pCT-NT-F2A, pCT-CT-F2A, pCT-CT-F2A-SED1SP, or pCT-CT-F2A-Prepro, were cultured in SGCAA media overnight, in an incubator-shaker at 20°C and 250 RPM.

Protocol tips
To induce cell surface protein expression, yeast cells containing pCTCON, pCT-NT-F2A, pCT-CT-F2A, pCT-CT-F2A-SED1SP, or pCT-CT-F2A-Prepro, were cultured in SGCAA media overnight, in an incubator-shaker at 20°C and 250 RPM.

Publication protocol

To induce cell surface protein expression, yeast cells containing pCTCON, pCT-NT-F2A, pCT-CT-F2A, pCT-CT-F2A-SED1SP, or pCT-CT-F2A-Prepro, were cultured in SGCAA media overnight, in an incubator-shaker at 20°C and 250 RPM. Subsequently, 5x106 cells were washed with PBS containing 0.1% BSA (PBS-BSA), and labeled with a chicken-anti-c-myc antibody (1:100 dilution) in 50 μl PBS-BSA for 20 minutes at room temperature. Secondary labeling was carried out using GAC633 (1:250 dilution) in 100 μL PBS-BSA for 12 minutes on ice. For simultaneous detection of cell surface expression and target binding, yeast cells expressing Sso7dhFc and Sso7dstrep were labeled with 500 nM biotinylated IgG and a 1:200 dilution of SA-PE, respectively, in a 50 μL reaction along with 1:100 dilution of chicken-anti-c-myc antibody. Secondary labeling was performed in a 100 μL reaction with a 1:250 dilution of GAC633 and 1:250 dilution SA-PE for Sso7dhFc, and 1:250 dilution GAC633 for Sso7dstrep. Samples were analyzed by a BD Accuri™ C6 flow cytometer (Franklin Lakes, NJ). Three independent replicate experiments were conducted, and fluorescence data for 50,000 cells was collected in each instance.

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