User:Jamie Nunziata/Notebook/Protease Research/2015/12/02

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Objective

To test our incubated fiber samples with various concentrations of a-chymotrypsin using Uv Vis (using Ocean Optics for a single data point), ICP Atomic Emission Spectrometer, and Conductivity.



Protocol

The incubated fiber samples fun yesterday were spun down to removed any fibers from solution. The supernatant was collected for the following:


Ocean Optics/UV Vis

Since the UV Vis lamp was down, we used Ocean Optics for our typical Uv Vis analysis by just collecting a single data point. 3mL of the supernatant from each of the 3 tubes for every protease concentration, along with the blank, added to the cuvette and absorbance was recordd using the Ocean Optics spectrometer. The cuvette was thoroughly cleaned after each trial to avoid contamination.



ICP

3mL of supernatant of each of the 13 samples was transferred to separate 15mL Falcon tubes. The samples were then tested by Dr. Hartins for gold ion concentration. The ICP was cleaned with DI water between each runs.



Conductivity

We measured the conductivity of each of our samples using a conductivity probe. 15mL of each of our samples were used to take these measurements.




Data

Figure 1: UV Vis/Ocean Optics Results

This table shows the results for the UV Vis performed by the Ocean Optics Spectrometer for the various a-chymotrypsin concentrations. As expected, the highest concentration of protease, 1000nm, had the highest absorbance, and as the concentration decreased the absorbance did as well


Figure 2: ICP Data Table


Image:JMN_ICP_RawDataTable_12_2.png

This table shows the raw data collected from the ICP for each of our samples. 100nM a-chymotrypsin sample 1 is the only unusual result because the gold ion concentration was significantly higher than the other two of the same concentration. This solution was purple, unlike the others, indicating that either a pipetting error occurred which put too much protease into the sample or the fibers were highly distrubed when transferring them from the jar to the falcon tubes, causing them to break up more and release more AuNPs into solution.


Figure 3: ICP graphical results



This figure shows the graphical results from the ICP data collected. Each data point is an average from the data collected at each protease concentration, with error bars to show the standard deviation. The overall trend is as expected, where the gold ion concentration increases as the concentration of a-chymotrypsin increases.


Figure 4: ICP graphical results for concentrations 0-100nM

This figure shows the ICP results without the 1000nM sample, to show clarity between the results in the 0-100nM range.


Figure 5: Conductivity Results

From this graph, it is clear that the conductivity does not necessarily depend on the protease or gold ion concentration, since the conductivity does not follow a specific trend unlike the data collected fro Ocean Optics and ICP



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