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Abstract：
Here, we present a protocol to quantify interactions among difficult-to-express proteins from Drosophila cells using the select western blot-free tagged-protein interaction (SWFTI) assay. We describe steps for plasmid design, cell plating, protein expression, and immunoprecipitation preparation. We then detail procedures for protein labeling, gel purification, and protein quantification. This protocol offers a fluorescence-based technique for rapid quantification of ectopically expressed proteins that are fused to SNAP and CLIP tags without the need for membrane transfer. For complete details on the use and execution of this protocol, please refer to Lin et al.1.
摘要：
这里,我们提出了一个方案,使用选择的westernblot-free标签蛋白相互作用(SWFTI)分析,量化果蝇细胞中难以表达的蛋白之间的相互作用.我们描述了质粒设计的步骤，细胞电镀，蛋白质表达，和免疫沉淀制备。然后我们详细说明蛋白质标记的程序，凝胶纯化，和蛋白质定量。该方案提供了一种基于荧光的技术，用于快速定量与SNAP和CLIP标签融合的异位表达的蛋白质，而无需膜转移。有关此协议的使用和执行的完整详细信息，PleaserefertoLinetal.1.
