Description

Laboratory Methods in Enzymology: Protein Part B brings together a number of core protocols concentrating on protein, carefully written and edited by experts.

Key Features

  • Indispensable tool for the researcher
  • Carefully written and edited by experts to contain step-by-step protocols
  • In this volume we have brought together a number of core protocols concentrating on protein

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

Contributors

Miscellaneous

Preface

Section I: Protein Protocols/Protein In Vitro Translation

Chapter One: In Vitro Synthesis of Proteins in Bacterial Extracts

1 Theory

2 Equipment

3 Materials

4 Protocol

5 Step 1 Grow and harvest E. coli for the S30 extract

6 Step 2 Preparation of the S30 extract

7 Step 3 Optimization of the coupled transcription and translation reaction

Chapter Two: Preparation of a Saccharomyces cerevisiae Cell-Free Extract for In Vitro Translation

1 Theory

2 Equipment

3 Materials

4 Protocol

5 Step 1 Preparation of Yeast Cell-Free Extract

6 Step 2 Cell-Free Translation

Section II: Protein Protocols/Protein In Vivo Binding Assays

Chapter Three: Yeast Two-Hybrid Screen

1 Theory

2 Equipment

3 Materials

4 Protocol

5 Step 1 Small-Scale Transformation of Yeast with pDBLeu-X

6 Step 2 Two-Hybrid Screen

7 Step 3 Confirmation of Positive Interactors

8 Step 4 Plasmid Rescue from Yeast

9 Step 5 Electroporation of E. coli with Yeast DNA and Identification of Positive Interactors

10 Step 6 Back-Transformation of Yeast and Further Confirmation of Interactions

Chapter Four: UV Cross-Linking of Interacting RNA and Protein in Cultured Cells

1 Theory

2 Equipment

3 Materials

4 Protocol

5 Step 1 UV Cross-Link RNA–Protein Complexes

6 Step 2 SDS Lysis of Cells

7 Step 3 Immunoprecipitation

8 Step 4 Proteinase K Treatment of RNA Samples

9 Step 5 RNA Analysis

10 Step 6 Protein Analysis

Chapter Five: Analysis of RNA–Protein Interactions by Cell Mixing

1 Theory

2 Equipment

3 Materials

4 Protocol

5 Step 1 Cell Mixing

6 Step 2 Cell Lysis

7 Step 3 Immunoprecipitation

8 Step 4 Protein

Details

No. of pages:
240
Language:
English
Copyright:
© 2014
Published:
Imprint:
Academic Press
eBook ISBN:
9780124201798
Print ISBN:
9780124201200

About the serial-volume-editor

Jon Lorsch

Affiliations and Expertise

Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA

Reviews

Laboratory Methods in Enzymology: Protein Part B brings together a number of core protocols concentrating on protein, carefully written and edited by experts.