BL483 Biochemical Techniques Lab - Week 8
Lab Report - What to Turn In for Lab Report 4
Prepare report with usual sections: Introduction, Methods, Results, Discussion
Native Page from Week 7
- Schematics of Native Gels for Protein and GOT Activity Stains
- Table of Relative Mobilities of Bands for Protein and Activity Stains
SDS-Page from Week 8
- Schematic of Protein Stain for Denaturing SDS Gel
- Mobilities of all bands in Gel for F-I, F-II, AS-II and CMC Fraction
- Plot of Std Protein MW (log) vs Relative Mobility on SDS-PAGE
- Table of Subunit MW estimates for all bands in CMC Fraction and Sigma GOT
- Native MW Estimate from Week 8
- Plot of log MW of Std Proteins vs Relative Elution (Ve/Vo)
- Estimate of Native MW of GOT
- Composition of GOT
- Use Subunit MW for CMC Fraction and explain how you decide which band on the SDS gel
represents the GOT Subunit
- Divide your Native GOT MW by Subunit MW for finding GOT composition
- Decide if GOT is a monomer, dimer or what for both your purified GOT and Sigma GOT
- Discussion
- Compare Native Page and SDS-PAGE - which is more highly resolving electrophoretic
method? Why do you think one is better than the other in resolution…What are the
advantages of each gel method?
- Compare PAGE results with Purification results from Lab Report 3 using the number of
protein bands found and specific activity of GOT, as well as total proteins in each
purification step
- Describe how you were able to decide which protein band is GOT in Native Gel and how
this helped to sort out the bands in SDS-PAGE gel.
- Compare SDS-PAGE results for your CMC fraction and Sigma GOT and describe any
differences found in the subunit MW of the two samples. Why do you think there would be
any difference between your CMC GOT and Sigma GOT?
- Compare the Composition of the Sigma GOT to your purified GOT fraction - do both GOT
samples have the same subunit composition?
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