Anti v5 fitc antibody

Saccharomyces cerevisiae CEN.PK 102-5B with different phenotypes (Additional file 7: Table S3) were cultured for 24 h for analysis. To detect the assembly efficiency of traditional dockerin and cohesin, secreted cellulases with dockerin in the supernatant were concentrated about tenfold and incubated with strains expressing scaffoldin ScafCipA3 at an OD₆₀₀ of 2, 5 and 15. The secreted cellulases incubated with ScafCipA3 expressing strains without concentration were used as controls. Cells were harvested and washed twice with phosphate-buffered saline solution (PBS, pH 7.0). The cells were then suspended in PBS containing 1 mg/mL bovine serum albumin to an OD₆₀₀ of 1.0. Monoclonal mouse antibodies including anti-V5-FITC antibody (Invitrogen) for scaffoldin, anti-DDDDK tag antibody (DyLight 488; Abcam, UK) for BGL1 and CBHI, and anti-Myc tag antibody (fluorescein isothiocyanate conjugated [FITC]; Abcam, UK) for CelA were used for immunofluorescence microscopy and FACS analyses. The antibodies were mixed with the cell suspension at 25 °C for 1 h with 1:500 dilution. Cells were harvested and washed twice with PBS after staining. Images were taken using immunofluorescence microscopy (Olympus, Japan) and flow cytometry analysis (FACS) was performed using a FACSCanto II system (BD FACSCanto II, USA).