User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2015/02/11: Difference between revisions
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The incorrect stock solution was used to make the protease solution. The concentration being tested is not 1 nM. It was calculated to be 1 µM of protease. This reaction will be rerun tomorrow with the proper enzyme concentration. | The incorrect stock solution was used to make the protease solution. The concentration being tested is not 1 nM. It was calculated to be 1 µM of protease. This reaction will be rerun tomorrow with the proper enzyme concentration. | ||
'''Proper Sample Prep''' | |||
#For 10 nM, take 69 µL of 0.4325 µL (10 µL stock, 990 µL buffer) protease, and dilute with 2.931 mL of buffer. | |||
#For 1 nM, take 6.9 µL of 0.4325 µL protease, and dilute with 2.993 mL of buffer. | |||
==Bradford Analysis of Samples from Yesterday== | ==Bradford Analysis of Samples from Yesterday== |
Revision as of 11:25, 11 February 2015
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In situ 1 nM Proteinase K Kinetics RunObjectives To run the 1 nM concentration of Proteinase K and track over time for kinetics workup. Protocol Take 0.046 mL of dilution from step 4 above in 1.954 mL of Tris/NaCl buffer for a final [proteinase K]final = 10 nM. From there, dilute by a factor of 10 to decrease the final concentration to 1 nM. Ocean Optics will be run at 37°C, scanning every 5 minutes for 6 hours. Results The incorrect stock solution was used to make the protease solution. The concentration being tested is not 1 nM. It was calculated to be 1 µM of protease. This reaction will be rerun tomorrow with the proper enzyme concentration. Proper Sample Prep
Bradford Analysis of Samples from YesterdayObjectives Bradford analysis of the samples extracted yesterday will be run. Protocol A 1 in 4 Bradford assay was made (1 mL Bradford in 3 mL of Tris/NaCl buffer). |