User:Daniel-Mario Larco/Notebook/AU Photosynthesis Lab/2014/07/30: Difference between revisions
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==SDS-PAGE== | ==SDS-PAGE== | ||
#We will run a gel with the solution and past solution to check purity | #We will run a gel with the solution and past solution to check purity | ||
#The gel will be set up in the following order | #The gel will be set up in the following order | ||
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##Excess Ruthenium solution | ##Excess Ruthenium solution | ||
##1st set of fractions from SP (first peak in Q-column 60 minute trial) | ##1st set of fractions from SP (first peak in Q-column 60 minute trial) | ||
##Empty | |||
##2nd set of fractions from SP (second peak in Q-column 60 minute trial) | ##2nd set of fractions from SP (second peak in Q-column 60 minute trial) | ||
##Injection 1 Q-column | ##Injection 1 Q-column | ||
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##Cleaning 2 " " | ##Cleaning 2 " " | ||
##Cleaning 3 " " | ##Cleaning 3 " " | ||
##Myoglobin and BSA ladder | ##Myoglobin and BSA ladder | ||
Revision as of 08:21, 30 July 2014
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Protein Purification
SDS-PAGE
Gel ElectrophoresisWe will be using a Bio-Rad Mini Protean system with pre-cast Mini Protean TGX gels. The manual for this system can be found here We will be running SDS-PAGE followed by gel development with Coomassie Blue staining. Below is a short description of how we will proceed. Please refer to the manual for more detailed instructions
Stock Solutions
-1mg BSA and 1mg Myoglobin in 1mL water -Solution was diluted to 1/10 to be used in gel
-14.4g Glycine, 1g SDS, 3g Tris in 100mL H2O -Solution was diluted to 1L with water
-40 Methanol, 10% Acetic Acid, 50% Water
-90% water, 10$ Acetic Acid and .0025% Commassie Brilliant Blue
-90% Water and 10% Acetic Acid |