User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2016/01/14

Biomaterials Design Lab

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Objective The referees of the microgel paper wanted me to determine if the protein hydrolyzes during the reaction for making hydrogels. So, I'm going to run 2 reaction (one with iron and one without iron) to see if the protein is degraded by the KOH and KNO3. today I am running the gel Description Using the BioRad MiniProtean electrophoresis setup Any kD Mini-Protean TGX gel Samples Hemoglobin in water (standard) 5 uL of sample (1.8 uM of Hemoglobin in water to match final concentration of hemoglobin from the micro gel sample prep) 5 uL of water 10 uL of 2X sample buffer Lane 5 Reaction with FeSO4 5 uL of sample 5 uL of water 10 uL of 2X sample buffer Lane 6 Reaction without FeSO4 5 uL of sample (1.8 uM of Hemoglobin in water to match final concentration of hemoglobin from the micro gel sample prep) 5 uL of water 10 uL of 2X sample buffer Lane 7 Running conditions Follow directions from Mini-Protean protocol Run voltage: 100 V Run time: 12: 05 pm to 12:56pm (dye band was about half-way down gel) Done this way so that I can observe any Hemoglobin fragments that may be made Gel Fixing Fixing solution 60mL of methanol, 15 mL of acetic acid, 75 mL water Remove gel from holder and place in fixing solution 1:02 pm to 1:32 pm Gel Staining Staining solution 37.5 mg coomassie blue, 15 mL of acetic acid, 135 mL water Stain from 1:35 until (should only be an hour. ran long due to meeting with the provost) Data Add data and results here... Notes This area is for any observations or conclusions that you would like to note. Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.