User:Jamie Nunziata/Notebook/Protease Research/2015/09/23

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Objective

The objective for today's lab work is to create variety lysozyme solutions from 1-10µg/mL and to analyze them using a Bradford assay in the UV Vis spectronomer


Procedure

We loosely based our procedure this experiment from the one Dr. Hartings had in his lab notebook.

First was creating a 5050µg/mL lysozyme solution by combining 0.1039g with 5mL of Tris buffer in a volumetric flask. Using the equation M1V1=M2V2, the amount Bradford Assay, lysozyme solution, protein assay reagent, and Tris buffer buffer needed for our various solutions. Those quantities can be found below:


Image:Nunziata_BradfordAssayConcentrations_9_23.png


These samples were then run through the Uv Vis spectrometer from 400-800nm for analysis.


Data

Figure 1: The results for the Bradford Assay at 530nm


Figure 2: Calibration Curve at 600nm

Image:Nunziata_Bradford_Calibration_Curve_9_23.png


Figure 3: Calibration Curve at 600nm wiithout Outlier

Image:Nunziata_Bradford_Calibration_Curve_without_Outlier_9_23.png

Since the calibration curve shows 10µg/mL to be on outlier. Without that value, our R2 value greatly increases


Overall, with the exception of the outlier, our results fit the desired trend where the lower the concentration of lysozyme, the lower the absorbance value (and vice versa)


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