Team:Washington/Protocols/Purified Enzyme Assay

From 2011.igem.org

(Difference between revisions)
(Created page with "{{Template:Team:Washington/Templates/Top}}")
Line 1: Line 1:
{{Template:Team:Washington/Templates/Top}}
{{Template:Team:Washington/Templates/Top}}
 +
 +
 +
=Whole Cell Lysate Assay=
 +
 +
<html>
 +
<script type="text/javascript">
 +
$(function() {
 +
  var mainimg = document.getElementById("mainimg");
 +
  var default_src = mainimg.src;
 +
  var restore_mainimg = function() {
 +
    mainimg.src = default_src;
 +
  };
 +
  $(document.getElementById("area_target")).hover(function() {
 +
    mainimg.src = "/wiki/images/1/1d/Main_graphic2_target.png";
 +
  }, restore_mainimg);
 +
  $(document.getElementById("area_secretion")).hover(function() {
 +
    mainimg.src = "/wiki/images/2/23/Main_graphic2_secretion.png";
 +
  }, restore_mainimg);
 +
  $(document.getElementById("area_display")).hover(function() {
 +
    mainimg.src = "/wiki/images/a/aa/Main_graphic2_display.png";
 +
  }, restore_mainimg);
 +
  $(document.getElementById("area_release")).hover(function() {
 +
    mainimg.src = "/wiki/images/e/ed/Main_graphic2_release.png";
 +
  }, restore_mainimg);
 +
});
 +
</script>
 +
</html>
 +
<!---------------------------------------PAGE CONTENT GOES BELOW THIS---------------------------------------->
 +
 +
 +
*'''Assay'''
 +
**Add 90uL of 5uM substrate (in NaAc pH 4.0 buffer) to each well of a black fluorescent microtiter assay plate
 +
**Start reaction by adding 0.0125mg/mL enzyme (try to avoid bubbles and pippette quickly, but accurately)
 +
***Use the P20 multichannel with a taped cardboard stopper to make sure you don't hit the pellet!
 +
**Monitor the reaction with the SpectraMax
 +
 +
 +
<!---------------------------------------PAGE CONTENT GOES ABOVE THIS---------------------------------------->
 +
<div style="text-align:center">
 +
 +
 +
'''&larr; [[Team:Washington/Protocols|Back to Protocols]]'''
 +
&nbsp; &nbsp; &nbsp;
 +
</div>

Revision as of 01:11, 23 September 2011


Whole Cell Lysate Assay


  • Assay
    • Add 90uL of 5uM substrate (in NaAc pH 4.0 buffer) to each well of a black fluorescent microtiter assay plate
    • Start reaction by adding 0.0125mg/mL enzyme (try to avoid bubbles and pippette quickly, but accurately)
      • Use the P20 multichannel with a taped cardboard stopper to make sure you don't hit the pellet!
    • Monitor the reaction with the SpectraMax



Back to Protocols