User:Jacob Esenther/Notebook/Chem 571/2014/11/12: Difference between revisions

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# UV Protein Analysis
# UV Protein Analysis
[[Image:Graphic_of_UV_of_Lysozyme_571.jpg|Plot of absorbance of Lysozyme in the UV range of 200-400nm]]
# Fluorescence Analysis
# Fluorescence Analysis


Revision as of 01:08, 14 November 2014

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Objective

  • Run all tests on the Lysozyme-CaCl2 dialysis
  • Make more lysozyme colloid solution
  • Set up a dialysis of Lysozyme-CaCl2 with new concentrations

Description

  1. Colloid Synthesis
    1. make 0.04621g HAuCl4 / 50mL H2O
    2. make 0.0448g Lysozyme / 25mL H2O
    3. make a 30:1 HAuCl4 Lysozyme solution
      • 0.5mL HAuCl4, 4.17mL Lysozyme, and 0.33mL 50mL H2O
    4. Heat in an oven over 4 hours at 80°C
  2. Its go time - 0.6g Lysozyme against 50mM, 5mM, 500μM, 50μM, 5μM CaCl2
    1. Bradford Analysis
      • 50μL sample
      • 200μL Bradford reagent
        • 4:1 DI Water to Bradford Assay
      • 750μL Tris-NaCl Buffer
        • 50mM Tris + 50mM NaCl
    2. UV Protein Analysis
      • 10:1 dilutions
    3. Fluorescence Analysis
      • 10:1 dilutions
    4. ISC of CaCl2
      • The probe was allowed to sit in solution before taking measurements, this seemed to yield better results. These measurements may need to be taken again to double check

Data

  1. Bradford Analysis

Graph plotting Absorbance of Lysozyme at differing Wavelengths

  1. UV Protein Analysis

Plot of absorbance of Lysozyme in the UV range of 200-400nm

  1. Fluorescence Analysis

Graph plotting Fluorescence at different Wavelength

  1. ISC of CaCl2
' ISC - CaCl2 Potential=26.344(Concentration)+40.946 ' ' '
Theoretical Concentration (mM) Potential (mV) log (concentration) Actual Concentration (mM)
Samples 50.000 89.7 1.850668084 70.90356695
5.000 65.6 0.93584877 8.62678094
0.500 46 0.191846341 1.555415207
0.050 32.1 -0.335788035 0.461542783
0.005 26.1 -0.563543881 0.273184541
Stocks 50.000 93.8 2.006301245 101.4614922
5.000 65.9 0.947236562 8.855978681
0.500 43.8 0.108335864 1.283322663
0.050 28.5 -0.472441543 0.336944566
0.005 15 -0.984892196 0.103539915

Notes

Always discuss methods between groups. It can only help you perfect your procedures.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.




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