Final answer:
Serum protein electrophoresis separates blood serum proteins by applying an electric current, causing them to migrate and form distinct bands based on their charge and molecular weight. Immunoelectrophoresis with PAGE further identifies proteins interactively with antisera, aiding in the discovery of antibody structure and the diagnosis of diseases like multiple myeloma.
Step-by-step explanation:
Serum protein electrophoresis is a laboratory technique used to separate proteins found in blood serum. The fundamental concept behind the procedure is that when an electric current is applied to the serum proteins in a gel, they will migrate at different rates based on their charge and molecular weight. Initially, plasma is rendered alkaline to ensure that proteins carry a negative charge and migrate towards the anode when electric current is applied. Serum proteins such as albumin, globulins, and fibrinogen separate into distinct bands on the gel which can be visualized using stains like Coomassie Brilliant Blue or silver stain. Immunoelectrophoresis, which includes a step of PAGE (polyacrylamide gel electrophoresis), is used to further differentiate proteins, especially in cases of elevated protein levels or specific diseases like multiple myeloma. Antisera are applied to the gel to form precipitin arcs with the proteins, identifying abnormal immunoglobulins. Two-dimensional gel electrophoresis further refines the process by separating proteins based on their isoelectric point (pI) and relative molecular mass (Mr), providing a detailed profile of thousands of proteins. Arne Tiselius laid the foundation for modern protein electrophoresis. His work allowed for the separation of proteins into albumin, alpha, beta, and gamma globulins, and set the stage for the discovery of antibody structure and the area of immunology that it would become. Modern applications of protein electrophoresis include diagnosing diseases such as multiple sclerosis and differentiating between types of immunoglobulins, particularly in cases like multiple myeloma where a distinct M spike is seen in the gamma-globulin region.