Preparation and Characterization of Dog Erythrocytic Membrane Antigen

Abstract

Dog erythrocytic membrane antigen plays a major role for determining blood group. Structural and molecular characterization of erythrocytic membrane antigen improves the production of blood typing antisera, to study the auto antibody production in canine autoimmune haemolyticanemia. The proteins in the lipid domain arranged from the inside of the erythrocyte to the outside. The integral membrane proteins include membrane protein 3 visible in coomassie Brilliant blue-stained polyacrylamide gels. The erythrocyte cytoskeleton consists of spectrin, ankyrin, actin, and protein 4.1 form a filamentous network under the lipid bilayer of erythroctic membrane. The cytoskeletal proteins interact with integral proteins and lipids of the bilayer forms network for maintaining membrane integrity. The external clustering of membrane protein 3 creates a recognition site for auto antibodies. The auto antibodies from the erythrocytes of dogs with acute immune haemolytic anemia have similar electrophoretic mobility to the human erythrocyte Rh related proteins and glycoproteins and able to immunoprecipitate membrane proteins that on SDS-PAGE and western blotting. The DEA 1 blood group system consists of antigens 1.1, 1.2, 1.3 which are most significant in terms of transfusion reactions and alloantibodies produced against DEA 3, 5 and 7 are not much clinical significance. Blood samples collected from registered donor dogs and RBC membrane were isolated by hypotonic freeze thaw method. The ghost membrane and cytoplasmic marker were identified in western blot.