Difference between revisions of "Part:BBa J70624"

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===Usage and Biology===
 
===Usage and Biology===
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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.
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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>
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<li>Yield/Purity: Fairly low yields</li>
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<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>
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<li>Affects Function?: Tag will likely not, but the low pH elution may</li>
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<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:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]