User:Omar Choudary/Notebook/Omar Choudary CHEM-571 AU-2011/2012/2011/11/16

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Contents

Objective

  • Perform gold nanoparticle(AuNP) synthesis experiment using 30 μM Bovine Serum Albumin (BSA), 4.17 mM Chloroauric Acid (HAuCl4), and H2O.
  • The following variables were tested:
  • Tube 1: Matched the Au/BSA ratio that yielded purple fibers.
  • Tube 2: Matched the Au/BSA ratio that yielded purple solution.
  • Tube 3: Test the effect of decreasing Gold.
  • Tube 4B: Test the effect of decreasing Gold.


  • Continue protein expression.

Description

  • AuNP synthesis:

Prepared four tubes to test different ratios of Au/BSA:

Tube 1: 0.1 mL BSA (30µM), 0.2 mL HAuCl4(4.17mM), 0.7 mL water. (Molar ratio of Au/BSA = 278)

Tube 2: 0.1 mL BSA (30µM), 0.143 mL HAuCl4 (4.17mM), 0.757 mL water. (Molar ratio of Au/BSA = 199)

Tube 3: 0.1 mL BSA (30µM), 0.115 mL HAuCl4 (4.17mM), 0.785 mL water. (Molar ratio of Au/BSA = 160)

Tube 4A: 0.1 mL BSA (30µM), excess of HAuCl4. (Molar ratio of Au/BSA = 0)

Tube 4B: 0.1 mL BSA (30µM), 0.086 mL HAuCl4 (4.17mM), 0.814 mL water. (Molar ratio of Au/BSA = 120)

Tubes kept at constant 80 degrees celsius in heatblock for 2 hours.


Image:OC-11-16-1.jpg


  • Protein expression:

The following had been added to each medium which had been prepared yesterday:

1 mL of Ampicillin (300 mg Ampicillin + 4 mL water)

1 mL of cell suspension (grown overnight)

1 mL of (380 mg IPTG (Isopropyl β-D-1-thiogalactopyranoside) + 4 mL water )

Placed the four mixtures in the shaker for several hours to facilitate cell proliferation

Centrifuged the four mixtures

Added medium to the cells and placed the mixtures into fridge set to -80 degrees celsius.

Data

Tube 1: Clear solution with yellow fibers.

Tube 2: Clear solution with purple fibers.

Tube 3: Homogeneous purple solution.

Tube 4A: Homogeneous yellow solution.

Tube 4B: Clear solution with yellow fibers.

  • Certain ranges of ratios of Au/BSA were more likely to yield clear solutions with purple protein fiber aggregates, lower ratios than those that yielded fibers were more likely to yield homogeneous purple solutions.
  • The results obtained from Tubes 1 and 2 were not expected compared to previous experiments. In addition to Au/BSA ratios, there are other factors that affect the formation of purple fibers or a homogeneous purple solution.

Notes

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