rotein analysis by gel electrophoresis  a).  Using the gel image provided to calculate the electrophoretic relative mobility as a ratio of the distance of protein migration to the distance of the tracking dye migration (See Appendix B).  If your dye front is not visible, measure the mobility relative to the bottom edge of the resolving gel.  Include a labeled print-out of your gel image with your report.  b).   Plot log MW of the protein markers and commercial myoglobin vs. mobility. Determine the molecular weight of myoglobin obtained after the final purification step (Sample E) from the equation of the line.  Submit a copy of your graph along with the gel image. c). Compare your sample E and commercial myoglobin with the ladder. Summary your results.   order of sample (left to right) is :  A  B  C  D  blank  E  blank  Commercial Myoglobin   Molecular Ladder

Human Anatomy & Physiology (11th Edition)
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Chapter1: The Human Body: An Orientation
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  1. Protein analysis by gel electrophoresis 

a).  Using the gel image provided to calculate the electrophoretic relative mobility as a ratio of the distance of protein migration to the distance of the tracking dye migration (See Appendix B).  If your dye front is not visible, measure the mobility relative to the bottom edge of the resolving gel.  Include a labeled print-out of your gel image with your report. 

b).   Plot log MW of the protein markers and commercial myoglobin vs. mobility. Determine the molecular weight of myoglobin obtained after the final purification step (Sample E) from the equation of the line.  Submit a copy of your graph along with the gel image.

c). Compare your sample E and commercial myoglobin with the ladder. Summary your results.

 

order of sample (left to right) is :  A  B  C  D  blank  E  blank  Commercial Myoglobin   Molecular Ladder

 

 

Appendix A:
Protein Molecular Weight Markers used in this lab:
kDa
|-116.0
B-galactosidase
66.2
Bovine serum albumin (BSA)
45.0
Ovalbumin
35.0
Lactate dehydrogenase
25.0
Restriction endonuclease Bsp981
18.4
14.4
B-lactoglobulin
Lysozyme
Commercial myoglobin, 16.0 kDa (not shown)
Appendix B: Example of how molecular weight of unknown protein
2
Myosin 200,000
Unknown
protein
B-Galactosidase 116,250
Glycogen phosphorylase b 97,400
Bovine serum albumin
66,200
Ovalbumin
45,000
Carbonic anhydrase
31,000
Soybean trypsin inhibitor
Lysozyme
21,500
14,400
Relative migration
м,
Unknown
standards protein
log M,
||| | |||
Transcribed Image Text:Appendix A: Protein Molecular Weight Markers used in this lab: kDa |-116.0 B-galactosidase 66.2 Bovine serum albumin (BSA) 45.0 Ovalbumin 35.0 Lactate dehydrogenase 25.0 Restriction endonuclease Bsp981 18.4 14.4 B-lactoglobulin Lysozyme Commercial myoglobin, 16.0 kDa (not shown) Appendix B: Example of how molecular weight of unknown protein 2 Myosin 200,000 Unknown protein B-Galactosidase 116,250 Glycogen phosphorylase b 97,400 Bovine serum albumin 66,200 Ovalbumin 45,000 Carbonic anhydrase 31,000 Soybean trypsin inhibitor Lysozyme 21,500 14,400 Relative migration м, Unknown standards protein log M, ||| | |||
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