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
- 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.
- 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.
- We synthesized AuNP fiber samples:
- We spun down 15 AuNP fiber samples at 300 RPM for 10 minutes. These samples had all been synthesized as 20mL aliquots.
- We pipetted off the supernatant. We collected this supernatant so that we could use it as our blank in our experiments tomorrow.
- We labeled our samples 1-15
- 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.
- 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
|
Project name
Main project page
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