User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/10/28
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To analyze the activity of proteinase k at 1 μM using fluorescence
The guide for analyzing a protease with fluorescence can be found here at Dr. Hartings notebook
To find the concentrations of peptide released at the various time intervals the samples and blanks had the area of their intensity integrated from 420 nm to 650 nm. The equation (Wavelength2 - Wavelength1)/2*0.5 was used and the sums of each interval totaled. The samples were then corrected for by subtracting the total area of the blank from the total area of the sample. The resulting intensities were then used to calculate the concentrations based off the calibration curve made on 10/07/15. The graph below shows the concentrations of the peptides as a function of time.
For the 1 μM fluorescence sample the 30 minute and 120 minute samples both had very high values for their blanks, which could explain why those values went negative. Also, 60 minutes is much lower than expected. Overall the graph does not appear to make any trend.