User:Andrew W Long/Notebook/CHEM-671 Research/2015/10/27

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Bradford Assay, 10nM

The instructions for the day can be found here

Previously synthesized fiber samples were prepared in the following way. First, 7 fiber samples were centrifuged at 300rpm for 10min, and the supernatant was removed. Stock of proteinase-K was suspended with 1mL of Tris buffer (50mM) with CaCl (10mM) at pH 8. The concentration of proteinase-K of the resulting solution was then determined to be 41868 μM. This was calculated using (0.00121g)(1mol/28900g)(1/0.001L). This solution was diluted in the following way to get a concentration of 0.238843 uL. (41868 nM)*(V1) = (10nM)*(1mL). The amount of buffer required to make the solution 1mL was determined to be 976.1uL to make 1mL of sample. Next, the 7 samples were incubated in a shaking hot water bath, as well as the 7 blanks. Once each sample had incubated for it's allotted time, it was pulled out of the bath, and centrifuged at 300rpm for 1min. 750mL was placed in a plastic cuvette, with 600mL dilute Bradford stock solution, and 1650uL buffer. The time periods were 10min, 15min, 30min, 45min, 1hr, 1.5hr, and 2hr. The samples were scanned from 400-800nm using the UV-Vis. The results from the experiment are shown below:

Bradford Assay of 10nM AuNP Fibers with Lysozyme Digestion, Wavelength vs. Absorbance at Various Times .png

AMS Bradford Assay of 10nM AuNP Fibers with Lysozyme Digestion, Absorbance at 600nm vs. Time.png