Western blot analysis of proteins expressed in yeast

Inma Quilis,Carmen Bano,Juan Carlos Igual

doi:10.1038/protex.2019.003

Abstract

Western blot analysis of proteins expressed in yeast

Highlights

This work is licensed under a Creative Commons Attribution 4.0 International
Collect the supernatant \(protein extract) – Do not touch the pelletand pass it into a new eppendorf
Equivalent amount of each protein sample is loaded into PAGE- gel based on this measurement

Summary

Introduction

- Resolving gel buffer: Tris-HCl 0.75 M pH 8.9, 0.2% SDS, 4 μM EDTA - Stacking gel buffer: 0.1 M Tris-phosphate pH 6.7, 0.2% SDS, 4 μM EDTA Electrophoresis buffer: 0.025 M Tris, 0.2 M glycine, 0.1% SDS - Transfer buffer: 0.025 M Tris, 0.2 M glycine, 0.1% SDS, 20% methanol - Western blot equipment: BIORAD electrophoresis and membrane transfer system - Refrigerator and freezer - Chemiluminescence reader ImageQuantTM LAS 4000mini biomolecular imager \(GE Healthcare).

Results

Conclusion