User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/10/25: Difference between revisions
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==Objective== | ==Objective== | ||
To run a protein gel to determine if the MBP-intein fusion has been expressed and purified. | To run a protein gel (SDS-PAGE) to determine if the MBP-intein fusion has been expressed and purified. | ||
==Description== | ==Description== | ||
# The stacking gel was prepared by miing the following in an Erlenmeyer flask: 17 mL of H<sub>2</sub>O, 4.15 mL of 30% arylamide, 3.15 mL of 1.0M Tris (pH 6.8), 0.25 mL of 10% SDS. | |||
# The following | # 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. | ||
# The stacking gel solution was the poured on top of the resolving gel and allowed to set for 20 minutes. | |||
# The four samples to be loaded into the the protein gel were prepared as shown in the box below. | |||
# Tube 1 was loaded into lane 1, tube 2 into lane 2, tube 3 into lane 3, and tube 4 into lane 4. | |||
# Electrophoresis was run for 40 minutes. | |||
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 | |||
==Data== | ==Data== | ||
[[Image:Group2.JPG]] | |||
[[Image:AllGels.JPG]] | |||
==Notes== | ==Notes== | ||
* Protein was present, but the sample was impure. Purification was not successful. | |||
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. | 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. |
Revision as of 15:06, 3 December 2011
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ObjectiveTo run a protein gel (SDS-PAGE) to determine if the MBP-intein fusion has been expressed and purified. Description
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 DataNotes
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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