Protocol to detect and quantify interactions between proteins expressed in Drosophila S2 cells

Abstract

Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Molecular Biology
• Molecular/Chemical Probes
• Protein Biochemistry
• Protein expression and purification