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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Engineering Saccharomyces cerevisiae for the production and secretion of Affibody molecules

Fig. 2

SDS-PAGE and western blot analysis of supernatant of B184k expressing different Affibody molecules. A SDS-PAGE of supernatant after 48 h of cultivation of B184k producing three different Affibody molecules (cropped). The lane marked with ZA sup contains supernatant of B184k expressing ZHER3_1-ABD, the lane marked with ZZA sup contains supernatant of B184k expressing ZHER3_1-ZHER3_1-ABD, the lane marked with ZAZ sup contains supernatant of B184k expressing ZHER3_1-ABD-ZHER3_1, the lane marked with ep contains supernatant of B184k with empty plasmid, the lane marked with ZA and st1 contains supernatant of ZHER3_1-ABD standard (12 kDa) dissolved in water, the lane marked with ZAZ and st2 contains supernatant of ZHER3_1-ABD-ZHER3_1 standard (18.9 kDa) dissolved in water. B Western blot against the Z-domain (cropped), the membrane was blotted with anti-Z-domain antibody (1:1000) followed by an anti-Mouse (1:5000) secondary antibody. C Western blot against the ABD (cropped), the membrane was blotted with anti-ABD antibody (3:1000) followed by an anti-Rabbit (1:5000) secondary antibody. D SDS-PAGE of supernatant of dissolved ZHER3_1-ABD in different solutions (cropped). The lane marked with media contains fresh SD2xSCAA media with (+) and without (−) ZHER3_1-ABD standard added, The lane marked with sup contains spent supernatant of a 24-h old culture of B184k in SD2xSCAA medium with (+) and without (−) ZHER3_1-ABD standard added, the lane marked with BSA contains water with 0.1% BSA with (+) and without (−) ZHER3_1-ABD standard added and the lane marked with H2O, + contains ZHER3_1-ABD standard in water. E Western blot against the ABD (cropped), the membrane was blotted with anti-ABD antibody (3:1000) followed by an anti-Rabbit (1:5000) secondary antibody

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