User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2014/02/18

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Biomaterials Design Lab: Spring 2014 Main project page
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Objective

  • To synthesize lysozyme-AuNPs.
  • Finish tests on 55:1 and 110:1 lysozyme-AuNPs as well as 100:1 BSA-AuNPs.
  • Image test results from last week.


Procedure

Creating Stock Solutions of Gold and Lysozyme

  • As seen below, each stock solution was created following the calculations in the table.
  • Each solution was created using a volumetric flask and deionized water.


Synthesizing Lysozyme-AuNPs

  • As seen below, each solutions was created using the following calculations.
  • We made ten tubes of each ratio.

Images

  • The 55:1 and 110:1 lysozyme samples consisted of pure fibers with variable added. However, the 100:1 BSA samples had no fibers before adding the variables.






Observations

OBSERVATIONS:

  • CHES: In the lysozyme AuNP fiber samples none of the fibers appear to have gone back into solution; however, there is a slight purple tinge to the solution. Similarly, for the BSA AuNP sample fiber formation wasn’t induced and the well appears identical to the control.
  • Phosphate Buffer: In the lysozyme AuNP fiber samples none of the fibers appear to have gone back into solution. Similarly, for the BSA AuNP sample fiber formation wasn’t induced and the well appears identical to the control.
  • HEPES: For the HEPES samples the Lysozyme AuNP fibers appear to have gone into solution slightly. The solutions appear to have more of a purple color than the other samples. Looking at the BSA well no fiber formation was seen and the shows no change from the control.
  • Citric Acid: In the lysozyme AuNP fiber samples none of the fibers appear to have gone back into solution. Looking at the BSA sample, citric acid appears to have caused the BSA AuNP to fall out of solution to form fibers. While solution still looks red there is a visible precipitate on the bottom of the well.
  • TRIS: Lysozyme AuNP fibers appear to have gone back into solution with application of TRIS. This is seen through the purple and pink coloration of the wells for these samples. The TRIS had no effect at all on the BSA AuNP samples.
  • MES: The lysozyme AuNP fiber samples appear to have gone back into solution slightly by the solution containing traces of a purple color. Adversely, in the BSA well MES was able to cause some of the AuNP’s to fall out of solution by creating a red aggregate.
  • EDTA: In the lysozyme AuNP fiber samples EDTA appears to have caused AuNP fibers to go back into solution slightly due to the increase in the purple color of the solution. EDTA appears to have had no effect of the BSA AuNP sample because the well looks identical to the control.
  • Chloroform: No effect was observed on the lysozyme fibers or on the BSA solution. There are two X’s on the spot plate because the solution evaporated to quickly for me to image.
  • EGTA, Acetone, DMF, and DMSO: All solutions had almost no effect of the lysozyme AuNP fibers. Only a slight pink tinge was observed in these samples, but physical fibers were still present in all wells. The BSA wells were not effected at all by the added variable.
  • 2, 2 Bipyridine: Had no effect on the Lysozyme fibers. This variable was able to cause fiber formation in the BSA well by creating very fine powdery looking fibers.
  • KCl, NaCl, MgCl2, and CaCl2: For all of these samples no change was observed in the lysozyme AuNP fiber samples. While it is difficult to see in these pictures. Very fine BSA fibers formed in all of the salt solution with the most extensive networks being found in the CaCl2 wells. The fibers cannot be seen in these pictures because the pictures were taken before the wells were allowed to settle long enough for the fibers to be easily seen.
  • SDS: Have very little effect on the lysozyme AuNP fibers. Overall all the fibers just appear slightly more red in color. The BSA AuNP wells has no visual change from the control.
  • 1, 4 Dioxane: No change was observed from the control.
  • HCl: It appears that the HCl broke down the fiber networks in the Lysozyme AuNP samples but didn’t cause them to go back into solution. This is noted because very few aggregates can be seen in the solutions. No effect on the BSA AuNP samples was noted with HCl.


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