Team:Washington/Notebook/Standard curve
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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 | ||
- | #Measure absorption at | + | #Measure absorption at 280nm to quantify protein amount |
- | # | + | #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
- Purify GFP-fused protein using traditional purification protocol
- Measure absorption at 280nm to quantify protein amount
- 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