User:Nicole Bonan/Notebook/Chem 571 Lab Notebook/2015/11/24

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Objective

The objective for today is to prepare AuNP fiber samples with varying concentrations of alpha-chymotrypsin to incubate overnight and use in experiments tomorrow.

Protocol

  1. We will be using the samples that we make today for ICP, Conductivity, and UV-Vis measurements tomorrow. We will make 20mL triplicates of AuNP fibers incubated for 24 hours in 1nM, 10nM, 100nM, and 1000nM (1µL) alpha-chymotrypsin. We'll also incubate three 20mL samples of AuNP fibers without the protease to serve as our zero-molar sample.
  2. From each 20mL sample, we'll use 5mL for ICP and 15mL for UV-Vis. We'll reuse the 15mL for Conductivity measurements as well.
  3. We synthesized AuNP fiber samples:
    1. We spun down 15 AuNP fiber samples at 300 RPM for 10 minutes. These samples had all been synthesized as 20mL aliquots.
    2. We pipetted off the supernatant. We collected this supernatant so that we could use it as our blank in our experiments tomorrow.
    3. We labeled our samples 1-15
    4. We then added 1mL of Tris buffer to two samples of alpha-chymotrypsin that we had measured out earlier in the semester. This brought the final concentration of the samples to 53.516µM and 57.03125µM.
    5. Next, we added the following concentrations of alpha-chymotrypsin and Tris buffer:


Sample # Molarity of alpha-Chymotrypsin Desired (nM) Volume of alpha-chymotrypsin Added (µL) Volume of Tris Buffer Added (µL)
1 0 0 20,000
2 0 0 20,000
3 0 0 20,000
4 1 3.51 (taken from part of the 57.03125µM stock that was diluted with Tris buffer in a 1:10 ratio) 19,996.49
5 1 3.51 (taken from part of the 57.03125µM stock that was diluted with Tris buffer in a 1:10 ratio) 19,996.49
6 1 3.51 (taken from part of the 57.03125µM stock that was diluted with Tris buffer in a 1:10 ratio) 19,996.49
7 10 3.51 (taken from the 57.03125µM stock) 19,996.49
8 10 3.51 (taken from the 57.03125µM stock) 19,996.49
9 10 3.51 (taken from the 57.03125µM stock) 19,996.49
10 100 35.1 (taken from the 57.03125µM stock) 19,964.9
11 100 35.1 (taken from the 57.03125µM stock) 19,964.9
12 100 35.1 (taken from the 57.03125µM stock) 19,964.9
13 1000 373.7 (taken from the 53.516µM stock) 19,626.3
14 1000 373.7 (taken from the 53.516µM stock) 19,626.3
15 1000 350.7 (taken from the 57.03125µM stock) 19,649.3


We then incubated all the samples (but not the blank) at 37 degrees Celsius for 24 hours.

Data

N/A



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