User:Lymperopoulos Loukas/Notebook/471 LAB/2015/09/30: Difference between revisions
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==Description== | ==Description== | ||
* Dr. Hartings left a procedure for us to use today in his lab [[User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2015/09/29 | notebook]]. | |||
*We are using the Pierce quantitative fluorometric peptide assay. (product link [https://www.thermofisher.com/order/catalog/product/23290 here]) | |||
# Perform a protease digest of lysozyme | |||
## Make a stock lysozyme solution (~ 1mg/mL) using phosphate buffer | |||
## Make your protease stock by adding 1mL to your pre-measured protease | |||
## Make a sample that has a final volume of 1mL and protease concentration of 1uM | |||
### Add the appropriate volume of protease sample to a clean eppendorf tube | |||
### Add a volume of lysozyme solution so that the final volume is 1mL | |||
## Make a blank sample (no lysozyme) | |||
### Add the appropriate volume of protease sample to a clean eppendorf tube | |||
### Add a volume of buffer solution so that the final volume is 1mL | |||
## Add both tubes to the 37C water bath for 1hr | |||
## Make standards for analysis | |||
### Standard A: 150uL of reaction sample | |||
### Standard B: 75uL of Standard A and 75uL of water | |||
### Standard C: 75uL of Standard B and 75uL of water | |||
### Standard D: 75uL of Standard C and 75uL of water | |||
### Standard E: 75uL of Standard D and 75uL of water | |||
### Standard F: 75uL of Standard E and 75uL of water | |||
### Standard G: 75uL of Standard F and 75uL of water | |||
### Standard H: 75uL of Standard G and 75uL of water | |||
## Make blanks for analysis | |||
==Data== | ==Data== |
Revision as of 00:13, 28 November 2015
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