User:Sara K. Mattson/Notebook/Chemistry 671/2016/09/14

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Fluorescence Purpose Use fluorescence to examine the rate at which myoglobin unfolds at pH5 Procedure Sample was made Myoglobin: 0.25mM Au, 12.5uM Myoglobin, 0.1mM HCl 3mL total Au 4.57mM (0.25mM)(3.0mL)=(XmL)(4.57mM) XmL= .164mL= 164uL Au Myoglobin 184uM (12.5uM)(3.0mL)=(XmL)(184uM) X=0.315mL= 315uL Myoglobin HCl 0.1mM (3.0mL)(0.01mM)=(1.0mM)(XmL) XmL=0.03mL= 30uL HCl Water Volume of water= 3000uL- 164uL-274uL-30uL= 2532uL Absorbance measurements were taken and saved every 5 minutes for 3 hours The instrument was set at 80 degrees Celsius The scan range parameters were start at 310nm and ending at 540nm The emission and exit slit were set at 10nm Solutions for UV-VIS preparation Stock Concentrations: Au 4.57mM Myoglobin 198mM Fructose 13.0mM 0 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 Myoglobinstock μL of Water 4 500 0 0 274 0 315 3917 5 50 0 0 274 0 315 4360 6 5 0 0 274 0 315 4405 7 0 0 0 274 0 315 4410 8 0 5 0 274 0 315 4405 9 0 50 0 274 0 315 4360 10 0 500 0 274 0 315 3910 11 0 0 5 274 0 315 4405 12 0 0 50 274 0 315 4630 0.0625 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 Myoglobinstock μL of Water 4 500 0 0 274 24 315 3910 5 50 0 0 274 24 315 4360 6 5 0 0 274 24 315 4405 7 0 0 0 274 24 315 4410 8 0 5 0 274 24 315 4405 9 0 50 0 274 24 315 4360 10 0 500 0 274 24 315 3910 11 0 0 5 274 24 315 4405 12 0 0 50 274 24 315 4360 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 Myoglobinstock μL of Water 4 500 0 0 274 48 315 3910 5 50 0 0 274 48 315 4360 6 5 0 0 274 48 315 4405 7 0 0 0 274 48 315 4410 8 0 5 0 274 48 315 4405 9 0 50 0 274 48 315 4360 10 0 500 0 274 48 315 3910 11 0 0 5 274 48 315 4405 12 0 0 50 274 48 315 4360 Results Wavelength Integrated Intensity First 15 Minutes 35-55 Minutes Notes The actual concentration of the myoglobin used in fluorescence was 184mM, but the calculations were done for 119mM.