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Laboratory methods in enzymology. Protein, Part D /
In this volume we have brought together a number of core protocols concentrating on Protein, carefully written and edited by experts.
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
London :
Elsevier Academic Press,
2015.
|
| Edición: | First edition. |
| Colección: | Methods in enzymology ;
v. 559. |
| Temas: | |
| Acceso en línea: | Texto completo Texto completo |
Tabla de Contenidos:
- Purification of His-tagged proteins
- Affinity purification of a recombinant protein expressed as a fusion with the maltose-binding protein (MBP) tag
- Immunoaffinity purification of proteins
- Affinity purification of protein complexes using TAP tags
- Strep-tagged protein purification
- Proteoloytic affinity tag cleavage
- Affinity pull-down of proteins using anti-FLAG M2 agarose beads
- Protein affinity prufication using intein-chitin binding protein tags
- Purification of GST-tagged proteins.
- Front Cover; Laboratory Methods in Enzymology: Protein Part D; Copyright; Contents; Contributors; Preface; Chapter One: Purification of His-Tagged Proteins; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Preparation; 4.2. Duration; 5. Protocol A: Purification of His-tagged Proteins Under Native Conditions; 5.1. Tip; 6. Step 1A Preparation of a Cleared E. coli Lysate Under Native Conditions; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 6.5. Tip; 6.6. Tip; 6.7. Tip; 7. Step 2A Batch Purification of His-tagged Proteins from E. coli Under Native Conditions.
- 7.1. Overview7.2. Duration; 7.3. Tip; 7.4. Tip; 7.5. Tip; 7.6. Tip; 8. Step 1B Preparation of a Cleared E. coli Lysate Under Denaturing Conditions; 8.1. Overview; 8.2. Duration; 8.3. Tip; 8.4. Tip; 8.5. Tip; 8.6. Tip; 8.7. Tip; 9. Step 2B Batch Purification of His-tagged Proteins from E. coli Under Denaturing Conditions; 9.1. Overview; 9.2. Duration; 9.3. Tip; 9.4. Tip; 9.5. Tip; 9.6. Tip; References; Referenced Literature; Source References; Referenced Protocols in Methods Navigator.
- Chapter Two: Affinity Purification of a Recombinant Protein Expressed as a Fusion with the Maltose-Binding Protein (MBP) Tag1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 5. Step 1 Equilibration of the Column; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 6. Step 2 Binding of the Protein Sample; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 7. Step 3 Removal of Unbound Proteins; 7.1. Overview; 7.2. Duration; 8. Step 4 Elution of the Bound Protein; 8.1. Overview; 8.2. Duration; 8.3. Tip; 8.4. Tip; 8.5. Tip; 8.6. Tip.
- 8.7. Tip8.8. Tip; References; Referenced Literature; Referenced Protocols in Methods Navigator; Chapter Three: Immunoaffinity Purification of Proteins; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 4.3. Tip; 4.4. Tip; 5. Step 1 Equilibrate Column; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 6. Step 2 Loading the Column; 6.1. Overview; 6.2. Duration; 6.3. Tip; 7. Step 3 Washing the Column; 7.1. Overview; 7.2. Duration; 7.3. Tip; 8. Step 4 Elution of the Protein from the Column; 8.1. Overview; 8.2. Duration; 8.3. Tip.
- 8.4. Tip8.5. Tip; 8.6. Tip; 9. Step 5 Column Regeneration and Storage; 9.1. Overview; 9.2. Duration; 9.3. Tip; References; Referenced Protocols in Methods Navigator; Chapter Four: Affinity Purification of Protein Complexes Using TAP Tags; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Preparation; 4.2. Duration; 5. Step 1 Preparation of Protein Lysates; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 5.5. Tip; 6. Step 2 Binding of the Protein A Tag to IgG-Sepharose; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 6.5. Tip; 6.6. Tip 7 Step 3 TEV Protease Cleave.