User:Andrew K Farag/Notebook/Andrewfarag/2013/09/04
Biomaterials Design Lab | <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> |
ObjectiveBecome familiar with the fluorescence spectrometer and with protein fluorescence. DescriptionFluorescence spectroscopy is another way to analyze molecular samples. InstructionsEach group will pick a different protein (bovine serum albumin, horseradish peroxidase, pepsin, adenosine deaminase, hemoglobin). You'll need to make 4 samples ranging from 10uM to 0.5uM. Remember, we don't have a lot of most of these proteins, so figure out how to make a stock solution using as little solid protein as possible. The fluorescence cuvette needs only 200uL, so you don't need to make more than that for each concentration that you're measuring.
NotesI made a stock solution of BSA today. Here is the info: BSA solution 0.0104g BSA (MW = 66776g/mol) in 0.0100mL water → 15.6μM DataUV-Vis Inosine Spectra is taking in lab, and the data of the calibration curve at 248 nm is shown in the graph below.
Calculations Using class DataAdenosine's average absorbance is 0.250606061, and its standard deviation is 0.152263534.
Extrapolation-UnknownThe wavelength of the Inosine unknown is 249 with absorbance of 0.05. After correcting the best fit line, Absorbance=11007*Concentration.Accordingly, absorbance of 0.05 would result a concentration of 4.54*10-6M Actual concentration is 2.37-5M. The percent difference error on the calculated concentration is 422.0%
|