Team:UC Davis/assay for measuring protein localization and activity

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measuring protein localization and activity:</big><br>
measuring protein localization and activity:</big><br>
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<div style="text-align: left;">&nbsp;&nbsp;&nbsp; &nbsp;&nbsp; <br>
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&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; We will utilize both the Western
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&nbsp;&nbsp;&nbsp;&nbsp; We are using western blotting as our assay to
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blotting technique and the plate reader to identify and quantify our proteins, GFP and Luciferase.<br>
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measure protein localization and activity, where we have tagged
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all of our constructs with a 6xHIS tag.&nbsp; This allows us to compare
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basal GFP or Luciferase activity inside (basal) and outside of the cell
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(medium).&nbsp; <br>
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<div style="text-align: left;">&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; We
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may also use fluorescence as an assay by comparing the
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brightness of the cell when it keeps it inside the cell, and when it
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secretes it into the medium.&nbsp; Both of our test genes, GFP and
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Luciferase are fluorescence proteins.<br>
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Revision as of 09:18, 15 October 2009

 
Assay for measuring protein localization and activity:
      
     We are using western blotting as our assay to measure protein localization and activity, where we have tagged all of our constructs with a 6xHIS tag.  This allows us to compare basal GFP or Luciferase activity inside (basal) and outside of the cell (medium). 
        We may also use fluorescence as an assay by comparing the brightness of the cell when it keeps it inside the cell, and when it secretes it into the medium.  Both of our test genes, GFP and Luciferase are fluorescence proteins.


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