Analysis of Signaling Pathways by Western Blotting and Immunoprecipitation

Silvia Marino; Aymen I Idris

doi:10.1007/978-1-4939-8997-3_7

Analysis of Signaling Pathways by Western Blotting and Immunoprecipitation

Methods Mol Biol. 2019:1914:131-143. doi: 10.1007/978-1-4939-8997-3_7.

Authors

Silvia Marino¹, Aymen I Idris²

Affiliations

¹ Division Hematology Oncology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA.
² Department of Oncology and Metabolism, University of Sheffield, Sheffield, UK. aymen.idris@sheffield.ac.uk.

PMID: 30729463
DOI: 10.1007/978-1-4939-8997-3_7

Abstract

This chapter describes the analysis of signaling pathways in bone cells by the use of western blotting and immunoprecipitation, including a step-by-step guide to cell culture techniques, cellular and subcellular fractionation, protein isolation, purification, measurement, electrophoretic transfer, and detection.

Keywords: Immunoprecipitation; Phosphorylation; Prenylation; Protein; Signaling; Western blotting.

MeSH terms

Animals
Blotting, Western / instrumentation
Blotting, Western / methods*
Bone and Bones / cytology*
Bone and Bones / metabolism
Cell Culture Techniques / instrumentation
Cell Culture Techniques / methods
Cells, Cultured
Electrophoresis / instrumentation
Electrophoresis / methods
Humans
Immunoprecipitation / instrumentation
Immunoprecipitation / methods*
Proteins / analysis*
Proteins / isolation & purification
Proteins / metabolism
Signal Transduction*

Substances

Proteins