Protocol for the isolation and immunoprecipitation of cell surface proteins from Drosophila melanogaster pupae

果蝇蛹细胞表面蛋白分离和免疫沉淀方案

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Abstract

Transmembrane proteins are notoriously difficult to isolate from tissue samples due to their hydrophobic nature and often low abundance. Here, we describe a protocol for the isolation, enrichment, and detection of the high-molecular-weight adhesion-type G protein-coupled receptor (aGPCR) member ADGRL/Latrophilin/Cirl from lipid-rich Drosophila melanogaster pupae. We detail steps to collect and homogenize pupae, enrich Cirl through immunoprecipitation, and visualize protein bands after SDS-PAGE and Western Blot. This protocol can be readily adapted to other tissues and (transmembrane) proteins. For complete details on the use and execution of this protocol, please refer to Bormann et al.(1).

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