Complete 5 pages APA formatted article: Western Blotting which is Used to Isolate Myosin Light Chain. What is more, western blotting helps scientists isolate specific proteins, e.g., Myosin light chains, from a composite mixture of proteins extracted from cells? Besides, this technique is dependent on three elements to realize this chore: (a) isolation based molecular size, (b) transfer to nitrocellulose, and (c) immunodetection to visualize. In this lab experiment, western blotting was used to isolate the myosin light chain from composite muscle cell extracts of closely and distantly related fish species.
Western blot is a technique used in scientific investigations to isolate as well as identify polypeptides. In this method, an assortment of polypeptides is resolved based on molecular weight, and hence by type, by gel electrophoresis. Consequently, the proteins are then relocated to a microporous membrane generating a band for each protein. The microporous membrane is then incubated with antibodies particular to the protein being studied. The unattached antibody is then washed off, leaving only the antibody attached to the protein investigated. The bound antibodies are then visualized by processing the film. Since the antibodies only bind to the protein being studied, only a single band should be detectable. Besides, the thickness of the band relates to the quantity of protein present. hence running a standard can demonstrate the protein present in the cell extract (Mahmood and Ping Chang, 2012).
In this research, protein gel electrophoresis (SDS-PAGE), as well as western blotting, was used to specifically identify a subunit of myosin light chain from the composite proteins that constitute the muscle tissues of diverse fish species.
In the experiment, the proteins in fish muscles were extracted through a lysis buffer’s lytic action to break open the muscle cells. The extraction buffer .contained the ionic agent sodium dodecyl sulfate (SDS) and a strong reducing agent DTT-dithiothreitol.