User:Benjamin Friedel/Notebook/CHEM 471/2015/10/14
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ObjectiveThe objective for today is to measure the UV absorbance of lysozyme AuNP fibers over various incubation times with alpha chymotrypsin using a bradford assay. ProtocolProtease Preparation
Incubation Tube Preparation (Samples and Blanks)
DataFigure 1 thumb|center|700px Figure 1 above shows absorbance data for both blanks and samples separately at 600nm along alpha-chymotrypsin incubation time. The absorbance data was corrected separately for both samples and blanks. First, the absorbance of the Bradford blank (bradford reagent and Tris buffer as specified above) was subtracted from the absorbance of each incubation timed sample and blank for each wavelength. Then, again individually, the isosbestic point was corrected for by subtracting the absorbance value at the isosbestic point of 535nm from each data point for each timed sample or blank respectively. This allowed for the absorbance values of all samples and blanks to be 0 at 535nm.
Figure 2 above shows the absorbance data of the AuNP samples over alpha-chymotrypsin incubation corrected for blanks. Blank correction was performed by subtracting the values of the Bradford blank and isosbestic point corrected blank data from the Bradford blank and isosbestic point corrected sample (AuNP fiber tube) data. The increase in absorbance as incubation time increases suggests that as alpha-chymotrypsin digests AuNP fibers samples, peptides and small fiber particles are released into solution. |