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

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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

  1. Using the BioRad MiniProtean electrophoresis setup
  2. Any kD Mini-Protean TGX gel
  3. Samples
    1. Hemoglobin in water (standard)
      1. 5 uL of sample (1.8 uM of Hemoglobin in water to match final concentration of hemoglobin from the micro gel sample prep)
      2. 5 uL of water
      3. 10 uL of 2X sample buffer
      4. Lane 5
    2. Reaction with FeSO4
      1. 5 uL of sample
      2. 5 uL of water
      3. 10 uL of 2X sample buffer
      4. Lane 6
    3. Reaction without FeSO4
      1. 5 uL of sample (1.8 uM of Hemoglobin in water to match final concentration of hemoglobin from the micro gel sample prep)
      2. 5 uL of water
      3. 10 uL of 2X sample buffer
      4. Lane 7
  4. Running conditions
    1. Follow directions from Mini-Protean protocol
      1. Run voltage: 100 V
      2. Run time: 12: 05 pm to 12:56pm (dye band was about half-way down gel)
        1. Done this way so that I can observe any Hemoglobin fragments that may be made
  5. Gel Fixing
    1. Fixing solution
      1. 60mL of methanol, 15 mL of acetic acid, 75 mL water
    2. Remove gel from holder and place in fixing solution
    3. 1:02 pm to 1:32 pm
  6. Gel Staining
    1. Staining solution
      1. 37.5 mg coomassie blue, 15 mL of acetic acid, 135 mL water
    2. 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.


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