User:Madeleine Y. Bee/Notebook/CHEM-571 2013F/2013/10/23: Difference between revisions
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== | ==October 23, 2013== | ||
Happy Mole Day! | |||
===Objective=== | ===Objective=== | ||
To measure the activity of pepsin in solution and pepsin-AuNPs in solution. | |||
===Description=== | ===Description=== | ||
*Make a 37°C water bath to hold at least 3 test tubes (same size as yesterday) | |||
====Pepsin Digest==== | |||
#Make 3 1mL dilutions for regular pepsin in 0.01N HCl at the following concentrations: | |||
#* 10μg/mL | |||
#* 15μg/mL | |||
#* 20μg/mL | |||
#Make 3 1mL dilutions for pepsin-AuNPs in 0.01N HCl at the following concentrations: | |||
#* 10μg/mL | |||
#* 15μg/mL | |||
#* 20μg/mL | |||
#Make blank for pepsin | |||
#*2.5mL hemoglobin solution in test tube in 37C bath | |||
#*5mL perchloric acid | |||
#*0.5mL pepsin dilution | |||
#*Remove from bath after 5 minutes | |||
#*Filter | |||
#*Take UV-vis spectrum | |||
#Make blank for pepsin-AuNPs | |||
#*2.5mL hemoglobin solution in test tube in 37C bath | |||
#*5mL perchloric acid | |||
#*0.5mL pepsin-AuNPs | |||
#*Remove from bath after 5 minutes | |||
#*Filter | |||
#*Take UV-vis spectrum | |||
#Make test for pepsin | |||
#*2.5mL hemoglobin solution in test tube in 37C bath | |||
#*0.5mL pepsin dilution | |||
#*Wait 10 minutes | |||
#*5mL perchloric acid | |||
#*Remove from bath after 5 minutes | |||
#*Filter | |||
#*Take UV-vis spectrum | |||
#Make test for pepsin-AuNPs | |||
#*2.5mL hemoglobin solution in test tube in 37C bath | |||
#*0.5mL pepsin-AuNPs | |||
#*Wait 10 minutes | |||
#*5mL perchloric acid | |||
#*Remove from bath after 5 minutes | |||
#*Filter | |||
#*Take UV-vis spectrum | |||
#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] | |||
====Pepsin-AuNP Analysis==== | |||
(picture) | |||
<u>Physical Characteristics (by ratio):</u><br.> | |||
#30- | |||
#50- | |||
#70- | |||
#90- | |||
#110- | |||
#130- | |||
#150- | |||
#170- | |||
#190- | |||
#210- | |||
<br.> | |||
*Centrifuge 1mL of each ratio of NPs for 30 minutes | |||
*Take UV-vis spectrum of each ratio sample | |||
*Save the sample | |||
===Data=== | ===Data=== |
Revision as of 08:44, 23 October 2013
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October 23, 2013Happy Mole Day! ObjectiveTo measure the activity of pepsin in solution and pepsin-AuNPs in solution. Description
Pepsin Digest
Pepsin-AuNP Analysis(picture) Physical Characteristics (by ratio):<br.>
<br.>
DataNotes |