User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2013/10/23: Difference between revisions
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#### Measure the absorbance at 280 --- This will be your A<sub>280</sub>(Test) | #### Measure the absorbance at 280 --- This will be your A<sub>280</sub>(Test) | ||
### Calculate the number of units/mg of active enzyme in each sample | ### Calculate the number of units/mg of active enzyme in each sample | ||
#### Units/mg = [A<sub>280</sub>(Test) - A<sub>280</sub>(Blank)]*1000/[10minutes*mg enzyme in reaction mixture] | #### Units/mg = [A<sub>280</sub>(Test) - A<sub>280</sub>(Blank)]*1000/[10minutes*mg enzyme in reaction mixture]</s> | ||
# Protein-AuNP analysis | # Protein-AuNP analysis | ||
## Take a picture and note physical description of each ratio | ## Take a picture and note physical description of each ratio | ||
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## Take a UV-Vis spectrum of each sample (Be considerate of other users here) | ## Take a UV-Vis spectrum of each sample (Be considerate of other users here) | ||
## After taking the spectrum, save the sample in a fresh eppy tube. | ## After taking the spectrum, save the sample in a fresh eppy tube. | ||
# Synthesis of HRP-AuNPs and Hemoglobin-AuNPs | |||
## Into test tube that will contain a total of 5mL add | |||
## Au solution such that the final concentration is 0.25mM | |||
## For Myoglobin make the following solutions to reflect an Au:Protein ratios | |||
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## For HRP make the following solutions to reflect an Au:Protein ratios | |||
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## Water up to 5mL | |||
==Data== | ==Data== | ||
<u>Stock Solutions</u> | |||
# Gold | |||
## 49.3 mg HAuCl<sub>4</sub>·3H<sub>2</sub>O in 50mL water ---> 2.50mM | |||
# HRP | |||
## 6.6 mg HRP in 100 mL water ---> 1.5μM | |||
# Myoglobin | |||
## 28.7mg myoglobin in 100mL water ---> 16.9μM | |||
==Notes== | ==Notes== |
Revision as of 09:43, 23 October 2013
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ObjectiveTo measure pepsin activity of 1) pepsin in solution and 2) pepsin-AuNPs in solution. We're going to try a slightly different protocol for this since the previous one see here didn't give the best results. The protocol for today is based off of this. We'll also be taking spectra of the pepsin-AuNP samples you made yesterday. DescriptionMake a 37C water bath that can hold at least 3 test tubes (same size as you used yesterday) at a time --- START THIS FIRST
DataStock Solutions
NotesThis area is for any observations or conclusions that you would like to note.
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