User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/11/11: Difference between revisions
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###Amount of Proteinase K solution needed for 1mL with 1μM concentration: M1*V1 = M2*V2 => (____ μM)*(V1) = (100 nM)*(1 mL) => V1 = 0.00__ mL | ###Amount of Proteinase K solution needed for 1mL with 1μM concentration: M1*V1 = M2*V2 => (____ μM)*(V1) = (100 nM)*(1 mL) => V1 = 0.00__ mL | ||
###Amount of Buffer solution need to get to 1mL: (1mL total)-(0.____ mL Protinase K solution) = 0.____ mL buffer | ###Amount of Buffer solution need to get to 1mL: (1mL total)-(0.____ mL Protinase K solution) = 0.____ mL buffer | ||
NEED TO GET DATA AS WELL AS DILUTION | NEED TO GET DATA AS WELL AS DILUTION | ||
#Incubating Samples | #Incubating Samples | ||
##____ μL Protinase K and ____ μL buffer were mixed in the 7 fiber tubes as well as 7 blank eppendorf tubes | ##____ μL Protinase K and ____ μL buffer were mixed in the 7 fiber tubes as well as 7 blank eppendorf tubes | ||
##Eppendorf tubes were incubated on a shaker in a 37°C water bath for | ##Eppendorf tubes were incubated on a shaker in a 37°C water bath for 15 minutes, 45 minutes, 75 minutes, 45 minutes, 105 minutes, 135 minutes, 1220 minutes and 1440 minutes for one of the fiber tubes and one of the blanks. | ||
#Running Samples | #Running Samples | ||
##Once the time for a sample was up the tube was centrifuged at 12000 rpm for 1 minute to collect the remaining fibers | ##Once the time for a sample was up the tube was centrifuged at 12000 rpm for 1 minute to collect the remaining fibers |
Revision as of 19:46, 24 November 2015
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ObjectiveTo analyze the activity of proteinase k at 1 nM using fluorescence DescriptionThe guide for analyzing a protease with fluorescence can be found here at Dr. Hartings notebook
NEED TO GET DATA AS WELL AS DILUTION
ResultsThe image below shows the intensity for the fluorescence of samples and blanks from 400 to 650 nm. 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. |