Western Blotting

Abstract
Western blotting is an analytic technique used around the world for many reasons; detecting infections, diseases, and particular proteins in a tissue. When western blotting, first the proteins are extracted and undergo Gel Electrophoresis, next it goes through elecroblotting, and finally detects the proteins. When the results after every step is completed, the membrane is analyzed based on where the bands are present in the gel. It has been told that actin and myosin is present in fish, making it tougher to chew in most cases. To test the theory of actin and myosin being present in fish making it tougher to chew was the purpose of this experiment. The proteins from Rainbow trout, Catfish, Pollock, Salmon, Tilapia, Mahi-mahi, Cod,
Western blotting is used to detect proteins in a tissue or extract. In a previous experiment a gentleman by the name of K. Nockler and many others were studying an infectious food borne disease called Human Trinchinellosis. Human Trinchinellosis is a disease caused by ingestion of infected pork, meat and other foods in which animals can be exposed to this parasite. He analyzed sera from pigs using a western blot, for the detection of anti-Trichinella-IgG. The results from his experiment using western blotting was that 144 of the sera from pigs were Trichinella-free and 159 pig sera were infected. His study and our experiments are similar, but instead of trying to find a disease or infection in this study, actin and myosin was the focus. The purpose of this study was to examine if actin and myosin were present in different types of fish proteins by observing the molecular weights of the proteins using kaleidoscope, by western blotting. We examined Rainbow Trout, Catfish, Pollock, Salmon, Tilapia, Mahi-mahi, Cod, Lake Erie Perch, KD and Actin/Myosin Standard proteins and analyzed where the bands of the proteins were present on the membrane and compared them to where the actin and myosin bands fell in the
The first stage is to extract the proteins. In order to do this you start by taking six 1.5 microcentrifuge tubes and six screw cap tubes and label with each of the different meat samples. Then, one inserts 50 μl of Laemmli sample buffer into to each microcentrifuge tube. A piece of each meat sample is then cut and transferred to its labeled microcentrifuge tube. The tube must be flicked 15 times and incubated at room temperature for 5 minutes to agitate the proteins. The 15 μl buffers from each centrifuge is transferred into its labeled screwcap microcentrifuge tube using a micropipette and then heated to 95°C to extract the