PrecipitationMBBS.ppt

Experiment 2 Precipitation Definition: Soluble Ag + Ab →Ag-Ab complex Classification: ① Precipitation reaction in solution ② Precipitation reaction in gels ③Immunoelectrophoresis Technique ④Immunoturbidimetry Precipitation Reactions In Gels Agar、Agarose Precipitation reactions between soluble antigens and antibodies can take place in semisolid media such as agar gels. When soluble antigen and antibodies are placed in wells cut in the gel, the reactants diffuse in the gel and form gradients of concentration, with the highest concentrations closest to the wells. Some where between the two wells, the reacting antigen and antibodies will be present at proportions that are optimal for formation of a precipitate. Double Agar Diffusion Test analysis of the properties of Ag Patterns of Identity Patterns of Nonidentity Patterns of Partial Identity: spur single agar diffusion test The wells contain antigen at different concentrations, while the antibodies are distributed uniformly in the agar gel. Thus, the precipitin line is replaced by a precipitin ring around the well. The distance the precipitin ring migrates from the center of the antigen well is directly proportional to the concentration of antigen in the well. Method Result Counter Immunoelectrophoresis pH8～8.6 buffer Ag： PI =5 MW：small Electric field force :cathode→anode，strong Electroosmosis: anode→cathode Electric field force Electroosmosis Direction of phoresis: cathode→anode Ab： PI =7 MW：large Electric field force :cathode→anode，weak Electroosmosis: anode→cathode Electric field force Electroosmosis anode→cathode — Cathode +Anode Ag Ab Result Rocket Immunoelectrophoresis Immunoelectrophoresis Immunoelectrophoresis involves separating a mixture of proteins in an electrical field (electrophoresis) followed by their detection with antibodies diffusing into the gel. a small drop of human serum is placed in a well cut in the center of a slide that is coated wi