How can we compare

and/or identify organisms?

 DNA
 Proteins
Muscle Proteins
 Muscles contain many proteins, some of which are
variable from organism to organism
 Actin and Myosin
– Form muscle fibers that allow muscles to contract and relax
– Most common muscle proteins in all animals
What is SDS-PAGE?

 SDS (Sodium dodecyl sulphate) is a

detergent used to denature proteins
and give them a negative charge
 PAGE: Polyacrylamide Gel
Electrophoresis

 It is a technique to separate proteins

by their molecular weight
Why do we need to
denature the proteins?
Polyacrylamide Gel
Cathode

Anode

Proteins separated by molecular weight

Actin and Myosin

Kaleidoscope standard
Myosin – blue
β-galactosidase – magenta
Bovine serum albumin – green
Carbonic anhydrase – violet
Soybean trypsin inhibitor – orange
Lysozyme – red
Aprotinin - blue
How can we interpret our
SDS-PAGE results?
 Percent commonality is a measure of
the similarity between the protein
composition of 2 samples

 How could this help us solve a

question of “identity” in the case of
different types of seafood?
Calculating Percent
Commonality
• Pairwise comparisons between species

# of proteins in common
100%
Total # of unique proteins
After running the gel

 Take the gels out from the gel box

 Cut the tape on the sides
 Label a petri dish with name & section
 Bring gel and dish over to the staining
area

 Clean up your area and don’t forget to

sign out
 Figure 1. SDS-PAGE standard curve for
proteins

molecular weight (kD) 1000

100

10

1
0 10 20 30 40 50

distance migrated (mm)

•Logarithmic scale graph

•Trendline is exponential, but on this type of graph it appears linear

Kaleidoscope Standard
MW (daltons)
on Tris HCl
gel

Protein Color

Myosin Blue 204,649

B-galactosidase Magenta 127,511

Bovine serum
albumin Green 85,130

Carbonic
anhydrase Violet 37,830
Soybean
trypsin
inhibitor Orange 30,906
Lysozyme Red 27,230
Aprotinin Blue 6,638