Team:Bilkent-UNAMBG/Demonstrate
All single transformed IDT BL21 cells were induced with IPTG. Western blot was performed following the heat release. All constructs were fused to Ag43. Therefore we were expecting to see bands around 46 kDa.IDT G3 (Rajpal 15 design with SCI57 a-b chains) and IDT G7 (SCI57 with its own linker a-b chains) were found out to be expressed. We performed immunocytochemistry (ICC) assay in order to observe the expression of our SCI constructs. Working principle of immunocytochemistry assay is to provide bacterial cultures that are producing His-Tag containing proteins with His-Tag binding antibodies and visualize the cells under fluorescence microscopy. With primary antibodies, we bind the His-Tag that is expressed with the SCI that is secreted to the cell surface. After primary antibodies, secondary antibodies are used in order to label the primary antibodies with fluorescence activity. Under fluorescence microscopy these cells are visualised. ICC Data for IDT-3 ICC Data for IDT-7
Testing Insulin Expression
Western Blot
ICC
