User:Klare Lazor/Notebook/Chem-496-001/2012/02/21

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Biomaterials Design Lab Main project page
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Objective

  • UV vis 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 ...... .00309L......3.09ml
    • 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

Image:Uv_bsa+dye_feb_21.png

Notes

  • spreading for the BSA +DYE was unusual, repeating experiment to make sure this is a repeating patten. In addition, uv of of just plain protein and fluorescence of plain protein will be taken to see if it contributes to this spread.
  • In addition, according to UV the concentration of gold present is around 1µM instead of 0.38µM which was calculated


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