The Use of the Western Blot Analysis to Identify Four Unknown Species of Fish

The Use of the Western Blot Analysis to Identify Four Unknown Species of Fish

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Protein extractions from unidentified fish samples were separated according to the molecular weights by SDS polyacrylamide gel electrophoresis. Since some of these proteins are shared between fishes, phylogenetic evaluation was reached. Western blot analysis was used to identify four unknown species of aquatic animals via comparison of actin/myosin bands. According to the results of this assay, the best estimate is that the unidentified aquatic animals are specimens of salmon, tilapia, cod, and shrimp, respectively.
Introduction
Western blot has been a revolutionary technique for identifying the expression of proteins within relative molecular biological samples that shared the same ancestor. Moreover, the sensitivity and specificity of the western blot (Immunoblotting) enables it a common technique for determining specific protein levels in clinical samples. Since the antibody specific to the antigen immunospecificity), it enables the target protein to be identified. Western blotting can produce quantitative data about that protein, which in this case the difference between bands in each of the protein samples. The western blot is an analytical technique used to detect specific proteins in the given sample of tissue homogenate or extract. The proteins are then transferred to a membrane (in this case, nitrocellulose), where they are stained with antibodies specific to the target protein [1] [2].
This experiment designed to analyze the phylogenetic of four unknown different fish muscles samples in order to predict the type of the aquatic creature. In this study, actin and myosin proteins were analyzed on account of their fairly high and constant expression in the animal muscles. Since the actin and myosin proteins are the two major...


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...and arthropod (shrimp/krill). Well number 7 has low protein actin and myosin and can be identified as shrimp, and wells 4, 5, 6 are identified as salmon, tilapia, and cod, respectively [3].
In figure 2, it is clear that the protein was transferred successfully from the gel to the membrane. The blot analysis was performed to detect if the protein was expressed.
The amount of protein present in each sample dilution in the blot corresponds with an absorbance value for that unknown protein extract. The antibody will bind to the protein (actin or myosin) and the Horse Raddish Peroxidase will catalyze the cleaving of the substrate to produce color upon antibody-protein binding. According to figure 3, 4, 5, 6 lanes are similar which confirm that they shared the same ancestor. However, lane 7 is slightly different which approves again it evolves from different ancestor.


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