Methods in enzymology. Volume 531, Protein, Part B : laboratory methods in enzymology / edited by John Lorsh.

Format:

Book

Contributor:

Lorsch, Jon.

Language:

English

Subjects (All):

Biotechnology--Methodology.

Biotechnology.

Physical Description:

1 online resource (240 p.)

Place of Publication:

San Diego, California : Academic Press, 2014.

Language Note:

English

Summary:

Laboratory Methods in Enzymology: Protein Part B brings together a number of core protocols concentrating on protein, carefully written and edited by experts.Indispensable tool for the researcher Carefully written and edited by experts to contain step-by-step protocolsIn this volume we have brought together a number of core protocols concentrating on protein

Contents:

Front Cover; Laboratory Methods in Enzymology: Protein Part B; Copyright; 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; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 4.3. Caution; 5. Step 1 Grow and harvest E. coli for the S30 extract; 5.1. Overview; 5.2. Duration; 5.3. Tip; 6. Step 2 Preparation of the S30 extract; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Caution; 6.5. Caution

7. Step 3 Optimization of the coupled transcription and translation reaction7.1. Overview; 7.2. Duration; 7.3. Tip; 7.4. Tip; References; Referenced Literature; Related Literature; Referenced Protocols in Methods Navigator; Chapter Two: Preparation of a Saccharomyces cerevisiae Cell-Free Extract for In Vitro Translation; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 5. Step 1 Preparation of Yeast Cell-Free Extract; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 6. Step 2 Cell-Free Translation; 6.1. Overview; 6.2. Caution

6.3. Duration6.4. Tip; 6.5. Tip; 6.6. Tip; 6.7. Tip; 6.8. Tip; Source References; Related Literature; Referenced Protocols in Methods Navigator; Section II: Protein Protocols/Protein In Vivo Binding Assays; Chapter Three: Yeast Two-Hybrid Screen; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & Buffers; 4. Protocol; 4.1. Preparation; 4.2. Construction of the DB fusion protein; 4.3. Construction of the AD fusion protein; 4.4. Testing for Self-activation of pDBLeu-X (and pPC86-Y); 4.5. Duration; 5. Step 1 Small-Scale Transformation of Yeast with pDBLeu-X; 5.1. Overview; 5.2. Duration

6. Step 2 Two-Hybrid Screen6.1. Overview; 6.2. Duration; 7. Step 3 Confirmation of Positive Interactors; 7.1. Overview; 7.2. Duration; 8. Step 4 Plasmid Rescue from Yeast; 8.1. Overview; 8.2. Duration; 9. Step 5 Electroporation of E. coli with Yeast DNA and Identification of Positive Interactors; 9.1. Overview; 9.2. Duration; 10. Step 6 Back-Transformation of Yeast and Further Confirmation of Interactions; 10.1. Overview; 10.2. Duration; References; Related Literature; Referenced Protocols in Methods Navigator; Chapter Four: UV Cross-Linking of Interacting RNA and Protein in Cultured Cells

1. Theory2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 4.3. Tip; 5. Step 1 UV Cross-Link RNA-Protein Complexes; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 5.5. Tip; 6. Step 2 SDS Lysis of Cells; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 6.5. Tip; 7. Step 3 Immunoprecipitation; 7.1. Overview; 7.2. Duration; 7.3. Preparation; 7.4. Tip; 7.5. Tip; 8. Step 4 Proteinase K Treatment of RNA Samples; 8.1. Overview; 8.2. Duration; 8.3. Tip; 9. Step 5 RNA Analysis; 9.1. Overview; 9.2. Duration; 9.3. Tip; 10. Step 6 Protein Analysis

10.1. Overview

Notes:

Description based upon print version of record.

Includes bibliographical references and indexes.

Description based on online resource; title from PDF title page (ebrary, viewed March 10, 2014).

ISBN:

9780124201798

0124201792