高美華醫(yī)學(xué)免疫學(xué)

1、Experiment 2Precipitation Definition:托 iSoluble Ag + Ab Ag-Ab complex Classification: Precipitation reaction in solution Precipitation reaction in gels Immunoelectrophoresis Technique ImmunoturbidimetryPrecipitation Reactions In Gels Agar Agarose Precipitation reactions between soluble antigens anda

2、ntibodies 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 reacti

3、ng antigen and antibodies will be present at proportions that areoptimal for formation of a precipitate.Double Agar Diffusion TestAg1.3Q0Ag1oAb-1相同/、Ag1 Q(QAg2,4Ab-1,2.4完全不同Ag1 OCAg1,20Ab-1,2部分相同釣、4雙向免疫擴(kuò)散試驗(yàn)Inwnurotogyanalysis of the properties of Ag Patterns of Identity Patterns of Nonidentity Patte

4、rns of Partial Identity: spurI single agar diffusion test5r 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

5、the center of the antigen well is directly proportional to the concentration of antigen in the well.MethodResult含抗體凝膠V沉淀環(huán)O抗原濃度單向免疫擴(kuò)散試驗(yàn)Counter ImmunoelectrophoresispHXbuffer Cathode+AnodeAg： PI =5 MW： smallElectric fieldforce :cathodeanode, strongElectroosmosis: anodecathodeElectric field force Elect

6、roosmosisDirection of phoresis:Ab： PI =7 MW： largeElectric fieldforce :cathodeanode, weakElectroosmosis:anodecathodeElectric field force anodeResultTTRocket Immunoelectrophoresis+含抗體凝膠o o o o孔中加待測(cè)抗原J 1 /1火箭免疫電泳ImmunologyImmunoelectrophoresisJr Immunoelectrophoresis involves separating a mixture of p

7、roteins in an electrical field (electrophoresis) followed by their detection with antibodies diffusing into the get a small drop of human serum is placed in a well cut in the center of a slide that is coated withagar get The serum is then subjected to electrophoresis, which separates the various com

8、ponents according to their mobilities in the electrical field. After electrophoresis a trough is cut along the side of the slides，and antibodies to human serum proteins are placed in the trough. The antibodies diffuse in the agar as do the separated serum prot&ns. 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