User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/10/25

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Objective

To run a protein gel to determine if the MBP-intein fusion has been expressed and purified.

Description

  1. The stacking gel was prepared by miing the following in an Erlenmeyer flask: 17 mL of H2O, 4.15 mL of 30% arylamide, 3.15 mL of 1.0M Tris (pH 6.8), 0.25 mL of 10% SDS.
  2. Once the pre-made resolving gel solution had set (after 20 minutes), 0.25 mL of 10% ammonium persulfate and 0.025 mL of TEMED were added to the stacking gel solution.
  3. The stacking gel solution was the poured on top of the resolving gel and allowed to set for 20 minutes.
  4. The four samples to be loaded into the the protein gel were prepared as follows:
     Tube 1 - 10 μL of ladder mixed with 4 μL of orange dye
     Tube 2 - 16 μL of protein from Peak 1 with 4 μL of orange dye
     Tube 3 - 16 μL of protein from Peak 2 with 4 μL of orange dye
     Tube 4 - 16 μL of protein from Peak 3 with 4 μL of orange dye
 5. Tube 1 was loaded into lane 1, tube 2 into lane 2, tube 3 into lane 3, and tube 4 into lane 4.

 6. Electrophoresis was run for 40 minutes.


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.



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