Western Blotting Essay

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Pages: 7

Western Blotting is used to Identify a Subunit of Myosin
Light Chain in the Proteins of All the Different Fish.
Chae Yee
Kent State University:
BSCI 30140 Lab

Abstract Western Blotting can be used to detect the Myosin actin light chain in different species of fish and is used to distinguish from different species based on variation, commonality, or evolutionary divergence. First, proteins are extracted from the tissue and loaded into a gel matrix. The matrix will separate the proteins according to size using an electric current. Proteins that are separated after are blotted from the gel and onto a paper membrane. An antibody is then added to the membrane paper and causes a colored reaction. Following the reaction, the results
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Smaller proteins move farther and faster through the gel than larger proteins. Figure 2 and 3 demonstrates that the different fish proteins migrated around the same distances and have similar molecular masses of myosin.

Figure 1: Picture of the gel after Western Blotting that is used to detect the myosin light chain. Molecular weights of the myosin light chain were blotted for comparison. The molecular weight standard is indicated in kilodaltons.

Row1 | Row 2 | Row 3 | Row 4 | Row 5 | Row6 | Row7 | Row8 | Row 9 | Tuna | Catfish | Pollock | Salmon | Tilapia | Mahi -Mahi | Cod | Lake Perch | A/M |

Figure 2: A standard curve used to measure and record the distances of the protein bands. The graph is used to determine the molecular masses of different myosins by comparing the mobility of unknown proteins with protein standards that are in the same gel. The molecular mass is plotted on the y-axis against the distances migrated for each protein on the x-axis. SAMPLE NAMETunaCatfishPollockSalmonTilapiaMahi-MahiCodLake PerchA/M | Estimated Mr from Blot252020202025252525 | Distance Migrated, mm75 mm78 mm78 mm80 mm78 mm72 mm74 mm74 mm74 mm | Calculated Mr1.3981.301.301.301.301.3981.3981.3981.398 |
Distance Migrated (mm)
Distance Migrated (mm) Protein Molecular Mass (kD) Protein Molecular Mass (kD)

Figure 3: A table showing the different fish samples and a