User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/10/28: Difference between revisions
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##7 AuNP fiber samples were spun at 300 RPM for 10 minutes | ##7 AuNP fiber samples were spun at 300 RPM for 10 minutes | ||
##The supernatant was removed but the fibers were retained | ##The supernatant was removed but the fibers were retained | ||
##Protinase K tube | ##Protinase K tube 1B was mixed with 1 mL of 50 mM pH 8 phosphate buffer | ||
###Protinase K concentration: ( | ###Protinase K concentration: (0.00166)*(1mol/28,900g)*(1/0.001L)= 0.0000574 M Protinase K | ||
###Amount of Proteinase K solution needed for 1mL with 1μM concentration: M1*V1 = M2*V2 => ( | ###Amount of Proteinase K solution needed for 1mL with 1μM concentration: M1*V1 = M2*V2 => (57.4 μM)*(V1) = (1 μM)*(1 mL) => V1 = 0.017 mL | ||
###Amount of Buffer solution need to get to 1mL: (1mL total)-(0. | ###Amount of Buffer solution need to get to 1mL: (1mL total)-(0.017 mL Protinase K solution) = 0.983 mL buffer | ||
#Incubating Samples | #Incubating Samples | ||
## | ##17 μL Protinase K and 983 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 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hrs, 1.5 hrs, and 2 hrs for one of the fiber tubes and one of the blanks. | ##Eppendorf tubes were incubated on a shaker in a 37°C water bath for 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hrs, 1.5 hrs, and 2 hrs for one of the fiber tubes and one of the blanks. | ||
#Running Samples | #Running Samples |
Revision as of 20:29, 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 μM using fluorescence DescriptionThe guide for analyzing a protease with fluorescence can be found here at Dr. Hartings notebook
ResultsThe image below shows the intensity for the fluorescence of samples and blanks from 400 to 650 nm. [[]] NEED TO RUN 10 MIN AND 15 MIN 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. [[]] NEED TO RUN 10 MIN AND 15 MIN |