User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/31

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Purpose

  • Au/Lysozyme samples were made with the same procedure used to make Au/BSA solutions.

Procedure

  • Lysozyme in powder form from chicken egg white obtained from Signma® is used to make the lysozyme stock solution.
  • 5mL of 70μM lysozyme stock solution was made in the following way:
    • 0.005g of lysozyme was weighted out
    • 0.005g of lysozyme was transferred to a 15ml falcon tube and mixed well with 5mL of distilled water.
    • See Melissa's Notebook for calculation that stated that 0.005g of lysozyme was needed for 5mL of 70μM of lysozyme stock solution.
  • The HAuCl4 used to make Au/Lysozyme samples was already made on 2012/09/05, and the concentration of the HAuCl4 was 10mM.
  • Au/Lysozyme samples was made with mole ratios listed below:
  20 - 30 - 40 - 50 - 60 - 70 - 80 - 100 - 120 - 130
  • Based off the calculations for Au/BSA samples, a few changes were made to calculate the amount of lysozyme and HAuCl4 needed for Au/Lysozyme samples.
  • The table below listed the initial conditions for both lysozyme and HAuCl4 both theoretically and actually:
- Lysozyme HAuCl4
Molecular Weight [g/mol] 14300 339.79
Theoretical Concentration [M] 7.00E-05 0.01
Volume of Water to add [L] 0.005 0.025
Moles Present [mol] 3.50E-07 0.00025
Theoretical Mass Needed [g] 0.005 0.0849
Actual Mass Weighted [g] 0.005 0.0892
Actual Concentration [M] 0.0007 10501
  • With the above calculation, the actual concentration of lysozyme came out to be 70μM and the actual concentration of HAuCl4 came out to be 10.501mM. The two concentration was used to calculate the amount of water, lysozyme, HAuCl4 needed to make the ratios listed above.
  • The table below listed the amount of water, lysozyme, HAuCl4 used for each Au/Lysozyme sample:
Au/Lysozyme ratio Lysozyme added [uL] HAuCl4 added [uL] Water added [uL] Final Concentration of Au[mM] Final Concentration of Lysozyme[uM]
20 114.3 15.2 7870.5 1 20
30 114.3 22.9 7862.8 1 30
40 114.3 30.5 7855.2 1 40
50 114.3 38.1 7847.6 1 50
60 114.3 45.7 7840 1 60
70 114.3 53.3 7832.4 1 70
80 114.3 60.9 7824.8 1 80
100 114.3 76.2 7809.5 1 100
120 114.3 91.4 7794.3 1 120
130 114.3 99 7786.7 1 130
  • The stock solutions were added with pipetman and non-sterile pipette tips. All samples were made in 10mL large test tubes. All samples were capped and wrapped in aluminum foil.
  • All samples were placed in incubator at 85°C for four hours then in room temperature to cool down.