User:Madeleine Y. Bee/Notebook/CHEM-571 2013F/2013/10/23

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(Pepsin-AuNP Analysis)
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===Description===
===Description===
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*Make a 37°C water bath to hold at least 3 test tubes (same size as yesterday)
 
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=====Pepsin Digest=====
 
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#Make 3 1mL dilutions for regular pepsin in 0.01N HCl at the following concentrations:
 
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#* 10μg/mL
 
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#* 15μg/mL
 
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#* 20μg/mL
 
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#Make 3 1mL dilutions for pepsin-AuNPs in 0.01N HCl at the following concentrations:
 
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#* 10μg/mL
 
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#* 15μg/mL
 
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#* 20μg/mL
 
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#Make blank for pepsin
 
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#*2.5mL hemoglobin solution in test tube in 37C bath
 
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#*5mL perchloric acid
 
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#*0.5mL pepsin dilution
 
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#*Remove from bath after 5 minutes
 
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#*Filter
 
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#*Take UV-vis spectrum
 
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#Make blank for pepsin-AuNPs
 
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#*2.5mL hemoglobin solution in test tube in 37C bath
 
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#*5mL perchloric acid
 
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#*0.5mL pepsin-AuNPs
 
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#*Remove from bath after 5 minutes
 
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#*Filter
 
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#*Take UV-vis spectrum
 
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#Make test for pepsin
 
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#*2.5mL hemoglobin solution in test tube in 37C bath
 
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#*0.5mL pepsin dilution
 
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#*Wait 10 minutes
 
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#*5mL perchloric acid
 
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#*Remove from bath after 5 minutes
 
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#*Filter
 
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#*Take UV-vis spectrum
 
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#Make test for pepsin-AuNPs
 
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#*2.5mL hemoglobin solution in test tube in 37C bath
 
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#*0.5mL pepsin-AuNPs
 
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#*Wait 10 minutes
 
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#*5mL perchloric acid
 
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#*Remove from bath after 5 minutes
 
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#*Filter
 
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#*Take UV-vis spectrum
 
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#Calculate the number of units/mg of active enzyme in each sample
 
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#*Units/mg = [A<sub>280</sub>(Test) - A<sub>280</sub>(Blank)]*1000/[10minutes*mg enzyme in reaction mixture]
 
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=====Pepsin-AuNP Analysis=====
=====Pepsin-AuNP Analysis=====
(picture)<br.>
(picture)<br.>

Revision as of 11:52, 23 October 2013

Experimental Biological Chemistry: Fall 2013 Main project page
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October 23, 2013

Happy Mole Day!

Objective

To measure the activity of pepsin in solution and pepsin-AuNPs in solution.

Description

Pepsin-AuNP Analysis

(picture)
Physical Characteristics (by ratio):

  1. 30-
  2. 50-
  3. 70-
  4. 90-
  5. 110-
  6. 130-
  7. 150-
  8. 170-
  9. 190-
  10. 210-


  • Centrifuge 1mL of each ratio of NPs for 30 minutes
  • Take UV-vis spectrum of each ratio sample
  • Save the sample

Data

Notes


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