Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis

Abstract

Summary: Here, we describe a protocol for extraction and tandem immunoprecipitation of the cytoplasmic and nuclear proteins from mouse testis for mass spectrometry. This protocol has been applied to knockin mice that express a meiotic protein of interest tagged with 3xFLAG-HA in the testis. The protocol is optimized for salt extraction of cytoplasmic and nuclear proteins from mouse frozen testes and thus can be used for a variety of proteins.For complete details on the use and execution of this protocol, please refer to Ishiguro et al. (2020) and Tanno et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Molecular Biology
• Protein Biochemistry
• Protein expression and purification
• Mass Spectrometry