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Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen
Paul Dowling1,2, Stephen Gargan1,2, Margit Zweyer3
1Department of Biology, Maynooth University, National University of Ireland, Maynooth W23F2H6, Co. Kildare, Ireland.
STAR Protocols
|December 30, 2020
Summary
This study details a protocol for comparing spleen proteomes in wild type and mdx-4cv mice, a model for dystrophinopathy. The methods cover sample preparation, mass spectrometry, and bioinformatic analysis for proteomic profiling.
Area of Science:
- Biochemistry
- Proteomics
- Animal Models
Background:
- Dystrophinopathy, such as Duchenne muscular dystrophy, involves progressive muscle degeneration.
- The mdx-4cv mouse is a model used to study dystrophin-related diseases.
- Comparative proteomic analysis can reveal molecular differences in disease states.
Purpose of the Study:
- To provide a detailed protocol for comparative proteomic profiling of spleen tissue.
- To analyze protein expression differences between wild type and mdx-4cv mice.
- To establish a methodology for mass spectrometry-based proteomic studies in mouse models.
Main Methods:
- Tissue homogenization and protein extraction from spleen samples.
- Protein concentration determination and enzymatic digestion.
- Bottom-up proteomic mass spectrometry and bioinformatic data analysis.
Main Results:
- The protocol enables detailed comparative proteomic analysis of spleen tissue.
- Identification of specific proteins and pathways affected in the mdx-4cv mouse model.
- Quantification of proteomic changes relevant to dystrophinopathy.
Conclusions:
- This protocol provides a robust framework for spleen proteomic studies in mouse models of disease.
- The methodology facilitates the discovery of biomarkers and therapeutic targets for dystrophinopathy.
- Comparative proteomics offers insights into the systemic effects of genetic muscle disorders.
