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Objective
- fluorescence of bsa plus dye
- remaking of nano particle dye solutions
Description
Concentration Curve
- 1mL of 0.05M phosphate buffer blank, 1µM of dye, 2 µM of dye, 5 µM of dye, 10 µM of dye, and unknown bsa+dye sample (30µL of solution + 1000µL of buffer)
Remake of nano particles + dye calculations
- note two different solutions of nano particles were used to make this solution
- 10:1 ratio Buffer: Acid has to be taken into consideration because we do not want the acid to affect the buffer and change the pH and not deprotonate the cysteine
- y= buffer x= gold/bsa
- 0.05 m/L Buffer * xL = 10 ( 0.000276 m/L * yL]
- x+Y = 0.003L
- y = 0.003L-x
- 0.05M/L (x) = 10 (0.000276mols/L x (0.003L-x))
- x= 1.5693 E-4 L, so buffer is 0.1569mL
- y=0.003L-1.569347E-4L, so 2.8432mL of au/bsa
- To determine how much dye
- BSA mols=0.004mM(2.843mL)= 1.137E-8 mols Au moles= 0.276mM (2.8342mL)= 7.8469E-7 moles
- BSA Mols * 66,000g/mol= 7.505E-4 grams of BSA x 0.06329 mass ratio of cysteins to bsa= 4.75 E-5 grams cysteine
- 4.75E-5 g of cysteine / 424.06 g/ mole cysteine= 1.12E-8 moles of Cysteine
- 1.12E-8 moles of cysteine x 4mol dye/1mol of cysteine = 4.4808 E-8 mols of dye (MW of dye) = 2.309E-3 grams of dyes
- Stock solution of dye
- 2.309E-3 * 10mL = 0.2309 g/L
- .2309 g/L * xL = 2.309E-5 g
- x= 1E-4L = 100µL
What contained in new solution... just multiplied solution by 4 to get larger volume..
- This time
- buffer 0.6276mL
- Au/BSA 11.3724 mL
- Dye 9.236E-5 g
- Total vol 12 mL
- solution was seen to be fluorescing more than last time. the solution was tinted slight orange/purple.
- Last time
- buffer 0.1569mL
- Au/ Bsa 2.8431 mL
- Dye 2.309E-5 g
- Total vol 3 mL
Data
Notes
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Biomaterials Design Lab
