Team:Washington/Notebook/Standard curve

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(→Generating a Standard curve for GFP concentration)
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====Generating a Standard curve for GFP concentration====
====Generating a Standard curve for GFP concentration====
#Purify GFP-fused protein using traditional purification protocol
#Purify GFP-fused protein using traditional purification protocol
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#Measure absorption at 280nM to quantify protein amount
+
#Measure absorption at 280nm to quantify protein amount
-
#Serialy dilute protein into plate and read fluorescence
+
#Serially dilute protein into plate and read fluorescence, excitation 485nm, emission 525nm
#Compare values of protein concentration to fluorescence to create a standard curve allowing for the comparison of fluorescence to protein level
#Compare values of protein concentration to fluorescence to create a standard curve allowing for the comparison of fluorescence to protein level

Revision as of 21:31, 18 October 2009

Generating a Standard curve for GFP concentration

  1. Purify GFP-fused protein using traditional purification protocol
  2. Measure absorption at 280nm to quantify protein amount
  3. Serially dilute protein into plate and read fluorescence, excitation 485nm, emission 525nm
  4. Compare values of protein concentration to fluorescence to create a standard curve allowing for the comparison of fluorescence to protein level
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