Difference between revisions of "Part:BBa J70624"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | The FLAG tag is a common epitope purification method. Unlike c-myc tagging, flag tagging yields high enough yields and purity to be used for purification as well as detection, but the same problems with a non-regenerative matrix and low pH elution still apply. | + | The FLAG tag is a common epitope purification/detection method. Unlike c-myc tagging, flag tagging yields high enough yields and purity to be used for purification as well as detection, but the same problems with a non-regenerative matrix and low pH elution still apply. Elution with EDTA is also possible. |
The tag can be cleaved off by enterokinase after the c-terminal lysine. | The tag can be cleaved off by enterokinase after the c-terminal lysine. | ||
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<li>Protein Sequence: DYKDDDDK </li> | <li>Protein Sequence: DYKDDDDK </li> | ||
<li>Matrix: Monoclonal Antibody</li> | <li>Matrix: Monoclonal Antibody</li> | ||
| − | <li>Yield/Purity: Fairly low yields</li> | + | <li>Yield/Purity: Fairly low yields, >90% purity</li> |
<li># Steps: One step</li> | <li># Steps: One step</li> | ||
<li>Fusion Location: N-terminus</li> | <li>Fusion Location: N-terminus</li> | ||
| − | <li>Affects Function?: Tag will likely not, but | + | <li>Affects Function?: Tag will likely not, but a low pH elution may.</li> |
</ul> | </ul> | ||
Latest revision as of 17:00, 9 August 2010
RFC12 FLAG Tag Head Domain
This is a RFC12 compatible version of BBa_T2004. It is a FLAG tag head domain meant for n-terminal fusions.
Usage and Biology
The FLAG tag is a common epitope purification/detection method. Unlike c-myc tagging, flag tagging yields high enough yields and purity to be used for purification as well as detection, but the same problems with a non-regenerative matrix and low pH elution still apply. Elution with EDTA is also possible.
The tag can be cleaved off by enterokinase after the c-terminal lysine.
Common tag information:
- Protein Sequence: DYKDDDDK
- Matrix: Monoclonal Antibody
- Yield/Purity: Fairly low yields, >90% purity
- # Steps: One step
- Fusion Location: N-terminus
- Affects Function?: Tag will likely not, but a low pH elution may.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]