User:Elaine Marie Robbins/Notebook/CHEM-496/2011/10/25: Difference between revisions

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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> SDS-PAGE</span>
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==Objective==


To run an SDS-PAGE to determine whether or not MBP-intein fusion proteins are being expressed cells comparable to the cells frozen on [[User:Elaine_Marie_Robbins/Notebook/CHEM-496/2011/09/28|09/28/11]].


==Description==


# Make solutions for a 12% discontinuous gel using the [http://ergo.berkeley.edu/be115/protean3.pdf BioRad Mini-PROTEAN instructions].
# Set resolving gel solution in gel holder with methanol on top to make the top of the gel flat.
# Allow gel to sit for 20 min.
# Remove methanol.
# Add stacking gel solution.
# Load gels into SDS-PAGE.
# Load ladder and protein solutions into gel.
# Run SDS-PAGE.
# Stain gels with Coomassie Brilliant Blue R250 solution.
## Dissolve 0.2555 g Coomassie Brilliant Blue R250 in 90 mL of methanol:water (1:1 v/v) and 10 mL of glacial acetic acid.
## Filter solution through a Whatman No. 1 filter.
# Place on shaker overnight.


==Objective==
==Data==
*Add your objective here
stuff [[User:Matt Hartings|Matt Hartings]] 21:41, 6 September 2011 (EDT)
 
==Description==
*Add description here
more stuff [[User:Matt Hartings|Matt Hartings]] 21:43, 6 September 2011 (EDT)


==Data==
Protein visibly traveled on gel after SDS-PAGE.
* Add data and results here...


==Notes==
==Notes==
This area is for any observations or conclusions that you would like to note.


Protein solutions used were from group 2:
* [[User:Samiye_Yaman/Notebook/Lab_571|Samiye Yaman]]
* [[User:Elizabeth_Ghias/Notebook/Experimental_Biological_Chemistry| Elizabeth Ghias]]


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.


[[Category:Course]]
[[Category:Course]]
[[Category:Miscellaneous]]
[[Category:Miscellaneous]]




Revision as of 09:29, 26 October 2011

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Objective

To run an SDS-PAGE to determine whether or not MBP-intein fusion proteins are being expressed cells comparable to the cells frozen on 09/28/11.

Description

  1. Make solutions for a 12% discontinuous gel using the BioRad Mini-PROTEAN instructions.
  2. Set resolving gel solution in gel holder with methanol on top to make the top of the gel flat.
  3. Allow gel to sit for 20 min.
  4. Remove methanol.
  5. Add stacking gel solution.
  6. Load gels into SDS-PAGE.
  7. Load ladder and protein solutions into gel.
  8. Run SDS-PAGE.
  9. Stain gels with Coomassie Brilliant Blue R250 solution.
    1. Dissolve 0.2555 g Coomassie Brilliant Blue R250 in 90 mL of methanol:water (1:1 v/v) and 10 mL of glacial acetic acid.
    2. Filter solution through a Whatman No. 1 filter.
  10. Place on shaker overnight.

Data

Protein visibly traveled on gel after SDS-PAGE.

Notes

Protein solutions used were from group 2:




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