高美華醫(yī)學(xué)免疫學(xué)precipitationmbbs課件

1、高美華醫(yī)學(xué)免疫學(xué)precipitationmbbs課件高美華醫(yī)學(xué)免疫學(xué)precipitationmbbs課件Precipitation Reactions In Gels Agar、AgarosePrecipitation 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

2、 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. 牛牛文庫文檔分享Precipitation Reactions In GelD

3、ouble Agar Diffusion Test 牛牛文庫文檔分享Double Agar Diffusion Testwww. analysis of the properties of AgPatterns of IdentityPatterns of NonidentityPatterns of Partial Identity: spur 牛牛文庫文檔分享 analysis of the properties o single agar diffusion testThe wells contain antigen at different concentrations, while

4、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. 牛牛文庫文檔分享 single ag

5、ar diffusion testTMethod 牛牛文庫文檔分享M 牛牛文庫文檔分享Result 牛牛文庫文檔分享R 牛牛文庫文檔分享 牛牛文庫文檔分享 牛牛文庫文檔分享 Counter Immunoelectrophoresis 牛牛文庫文檔分享 Counter Immunoelectrophoresis pH88.6 bufferAg： PI =5 MW：small Electric field force :cathodeanode，strongElectroosmosis: anodecathodeElectric field force ElectroosmosisDirectio

6、n of phoresis: cathodeanodeAb： PI =7 MW：largeElectric field force :cathodeanode，weakElectroosmosis: anodecathodeElectric field force Electroosmosisanodecathode Cathode +Anode AgAb 牛牛文庫文檔分享 pH88.6 bufferAg： PI =5 Result 牛牛文庫文檔分享R 牛牛文庫文檔分享 牛牛文庫文檔分享 牛牛文庫文檔分享Rocket Immunoelectrophoresis 牛牛文庫文檔分享Rocket I

7、mmunoelectrophoresis w ImmunoelectrophoresisImmunoelectrophoresis involves separating a mixture of proteins in an electrical field (electrophoresis) followed by their detection with antibodies diffusing into the gel. 牛牛文庫文檔分享 ImmunoelectrophoresisImmunoela small drop of human serum is placed in a we

8、ll cut in the center of a slide that is coated with agar gel. The serum is then subjected to electrophoresis, which separates the various components according to their mobilities in the electrical field. After electrophoresis, a trough is cut along the side of the slides, and antibodies to human ser

9、um proteins are placed in the trough. 牛牛文庫文檔分享a small drop of human serum isThe antibodies diffuse in the agar, as do the separated serum proteins. At an optimal antigen:antibody ratio for each antigen and its corresponding antibodies, they form precipitin lines. Comparison of the pattern and intensity of lines of normal human serum with the patterns and intensity of lines obtained with sera of patients may reveal an absence, overabundance, or