| Section
2A: |
About
the Molecular Tagging Procedures |
2.2
|
| Section
2B: |
Addition
of an Epitope Tag Sequence to a Target Gene by PCR |
2.3
|
|
Critical
factors for success: Designing PCR primers for epitope tagging |
2.3
|
|
Getting
started: Procedures for epitope tagging by PCR |
2.6
|
|
Troubleshooting
the PCR procedure |
2.8 |
|
Specific
application of PCR procedures |
2.8 |
| Section
2C: |
Addition
of an Epitope Tag Sequence to a Target Gene via an Adaptor -Duplex |
2.10
|
|
Critical
factors for success: Designing an adaptor-duplex using the epitope tag sequence |
2.10
|
|
Getting
started: Procedures for adding a c-myc, HA, or VSV-G adaptor-duplex to the
N-terminal coding region of a target gene on a prokaryotic expression vector |
2.11
|
| Section
2D: |
Strategies
for Construction of an Expression Vector Containing an Epitope Tag Sequence
|
2.13 |
|
What
an epitope-tagging vector should contain |
2.13
|
|
Existing
epitope-tagging vectors |
2.14 |
| Section
4A: |
Overview
of Immunoaffinity Purification Methods |
4.3
|
| Section
4B: |
Critical
Factors for Successful Analysis and Purification of Tagged Proteins |
4.4
|
|
Preventing
proteolysis during sample purification |
4.4
|
|
Preparation
of cell lysate |
4.4 |
| Section
4C: |
Purification
by Affinity Chromatography |
4.6
|
|
Overview
of technique |
4.6
|
|
Getting
started: Procedures for affinity chromatography |
4.6
|
|
Troubleshooting
affinity chromatrgraphy |
4.12
|
| Section
4D: |
Purification
by Immunoprecipitation |
4.15
|
|
Overview
of technique |
4.15
|
|
Critical
factors for successful immunoprecipitation |
4.15
|
|
Getting
started: Procedures for immunoprecipitation |
4.16
|
|
Tips
on using other tag-specific antibodies for immunoprecipitation |
4.20
|
|
Troubleshooting
the immunoprecipitation procedure |
4.21
|
|
Specific
applications |
4.22
|
