《医学免疫学：Enzyme Linked Immunosorbent Assay》由会员分享，可在线阅读，更多相关《医学免疫学：Enzyme Linked Immunosorbent Assay（12页珍藏版）》请在金锄头文库上搜索。
1、ELISA Enzyme Linked Immunosorbent AssayJUN CHENDept. of microbiology and immunologyHuman anti-bodyanimalinjectAnti-antibodyPrimary antibodySecondary antibodyantigenSubstratePrimary antibodyEnzymeSecondary antibodyantigens Coloured productPrincipleSolid supportNo-colourEnzymeSecondary antibodyObjecti
2、ve To detect HBsAb in samples Master the principle of ELISAMaterials1. Diagnostic Kit for Antibody to Hepatitis B Surface Antigen（ELISA） 2. pipette, tips ,incubator, mark pencil and so on . 1、add samples (primary antibody) + - 1 2 3 4 5 6 Using a clean pipette, add 50l of different sample to each we
3、ll 16. Positive control ：1 drop Negative control : 1 dropMethods6 persons/group, 1 well/ person2、add secondary antibody (enzyme) + - 1 2 3 4 5 6add 1 drops of secondary antibody (anti-human IgG) to each well.enzymesecondary antibody (enzyme) mix, leave on bench for 30 minutes, 37 .3、Wash wells of ex
4、cess antibodies after 30 min, empty out contents of wells into waste container. Using pipette, fill wells with wash buffer then empty out. Tap wells upside down on paper towel. Repeat five times, making sure no liquid remains after the last wash. + -?samples4、add substrate for enzymeadd 1drop of sub
5、strate A and B to each well. Observe colour development (blue product formed) and compare with positive and negative control. + - mix, leave on bench for 10 minutes, 37 .4. Add substrate for enzyme1. Add primary antibody3. Wash with buffer2. Add secondary antibody5. Observe colour developmentResults
6、 + -Observe colour change (blue product formed) and compare with positive and negative control. Intensity of colour indicates amount of HBsAb present in sample. Conclusions?Exam (one by one)1.Preparation of agar plate (show to teacher)2.Punch wells on the agar (on your seat)3. Add samples (show to teacher)
