User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2013/09/24: Difference between revisions
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# 6.0mL 10% Sodium Dodecyl Sulfate (SDS) ---> 1g SDS in 10mL water | # 6.0mL 10% Sodium Dodecyl Sulfate (SDS) ---> 1g SDS in 10mL water | ||
# H<sub>2</sub>O to make 30mL total | # H<sub>2</sub>O to make 30mL total | ||
The class ran out of some solutions and Karlena made new: | |||
<u>Hemoglobin</u> | |||
250.1 mg hemoglobin in 50mL water | |||
and | |||
<u>Glycine HCl buffer</u> | |||
0.37910g glycine in 100mL water and adjust the pH to 3 ---> 50.5mM Glycine-HCl buffer pH 3 | |||
==Notes== | ==Notes== |
Revision as of 11:48, 24 September 2013
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ObjectiveTo observe the catalytic activity of pepsin (in cleaving peptide bonds in hemoglobin) in the presence and absence of pepstatin. This data will be compared with data we take in a later lab on pepsin-AuNPs. DescriptionThe experimental details (pepsin cleavage of hemoglobin) are similar to this reference. We will allow the pepsin cleavage of hemoglobin to proceed for two hours with aliquots taken and analyzed every 1/2 hour. We will also run parallel reactions with different concentrations of pepstatin (pepsin inhibitor) present. We will save our aliquots for analysis tomorrow using SDS-PAGE.
DataMaking the solutions Glycine-HCl Buffer pH 3
50 mL of 5 mg/mL Hemoglobin
Pepstatin Pepstatin is not very soluble in water. Solubilize in methanol as per wikipedia. 4.0mg pepstatin in 5mL methanol ---> 1.2mM pepstatin SDS-PAGE Sample Buffer (No DTT)
The class ran out of some solutions and Karlena made new: Hemoglobin 250.1 mg hemoglobin in 50mL water and Glycine HCl buffer 0.37910g glycine in 100mL water and adjust the pH to 3 ---> 50.5mM Glycine-HCl buffer pH 3 NotesThis area is for any observations or conclusions that you would like to note.
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