User:Patrick Hampson/Notebook/chem471/2016/09/07

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Fluorescence 201697 Protocol Starting the Peltier controller Make sure that the ribbon cord connects the peltier controller and the cuvette holder Fill the chiller bucket with water Place the aquarium pump (with tubes attached to the Peltier heat exchanger) into the bucket and plug in the pump so that water flows Turn on the peltier controller (switch in the back of the instrument) Use the arrow buttons to set the temperature. Turning off the Peltier controller Turn the controller power off Unplug the aquarium pump Dump the water from the bucket into the sink Sample Preparations stock solutions BSA 25.33 mg of myoglobin with water in 10.0 mL volumetric flask MW = 66776 g/mol concentration = 37.93 μM samples BSA Fluorescence 3 mL total 0.25 mM Au 3.125 μM BSA HCl or NaOH appropriate for your pH water BSA for UV-Vis during next session 5 mL total 0.25 mM Au 3.125 μM BSA HCl or NaOH appropriate for your pH Appropriate amount of fructose water Fluorescence data This graph was comprised from fluorescent scans taken once every five minutes. There is a steady increase in the peaks of the data over the course of time. After the initial datapoint, there is a drastic increase in emissions. This increase is not constant, as the rest of the data collected throughout the duration of the experiment were relatively similar. The datapoints appeared to form a plateau, however upon closer inspection, there is a slight increase in the emissions as time goes on. This graph displays the integrated fluorescence over time. The data resembles a minor parabola for the first 50 minutes of the experiment. The data then flattens out to just below 5,000 until it reaches 100 minutes. At 100 minutes there is a severe spike in the data, and integrated fluorescence increases to over 20,000. There is a steady linear increase in the integrated fluorescence for the remainder of the experiment. UV-Vis 5 mL total 0.25 mM Au 3.125 μM BSA HCl or NaOH appropriate for your pH Appropriate amount of fructose water stock solutions The NaOH and HCl samples were made exactly as designed. AuCl3 mass = 38.5 mg 25.0 mL volumetric flask concentration = 5.08 mM Fructose mass = 23.48 mg 10.0 mL volumetric flask concentration = 13.0 mM Bovine Serum Albumin mass = 25.33 mg 10.0 mL volumetric flask concentration = 37.93 mM 0.125 mM Fructose pH μL of 1 mM HClstock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AuCl3stock μL of Fructosestock μL of BSAstock μL of Water 4 500 0 0 246 48 408 3798 5 50 0 0 246 48 408 4248 6 5 0 0 246 48 408 4293 7 0 0 0 246 48 408 4298 8 0 5 0 246 48 408 4293 9 0 50 0 246 48 408 4248 10 0 500 0 246 48 408 3798 11 0 0 5 246 48 408 4293 12 0 0 50 246 48 408 4248 0.25 mM Fructose pH μL of 1 mM HClstock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AuCl3stock μL of Fructosestock μL of BSAstock μL of Water 4 500 0 0 246 96 408 3750 5 50 0 0 246 96 408 4200 6 5 0 0 246 96 408 4245 7 0 0 0 246 96 408 4250 8 0 5 0 246 96 408 4245 9 0 50 0 246 96 408 4200 10 0 500 0 246 96 408 3570 11 0 0 5 246 96 408 4245 12 0 0 50 246 96 408 4200 Solutions before being heated in the oven (low reactivity): Matt Hartings 14:02, 9 September 2016 (EDT): I remade your samples because you got strange results. Here's my description