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Experiment ten

Experiment ten. To detect the HBsAg with ELISA. ELISA Enzyme linked immunosorbent assay is a normal and simple means to detect the Ag and Ab of HBV and other pathogens. ELISA. ( E nzyme- L inked I mmuno s orbent A ssay).

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Experiment ten

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  1. Experiment ten To detect the HBsAg with ELISA

  2. ELISA Enzyme linked immunosorbent assay is a normal and simple means to detect the Ag and Ab of HBV and other pathogens

  3. ELISA (Enzyme-Linked Immunosorbent Assay)

  4. ELISA for Antibody Detection(Enzyme-Linked Immunosorbent Assay) • Capture of antigens in microplate • Washes • Block unbound sites in wells • Washes • Add antibody serum to wells • Washs • Add 2nd antibody (attach enzyme) • Washes • Color development • Ex: HIV test

  5. Sandwich ELISA for Antigen Detection(Enzyme-Linked Immunosorbent Assay) • Capture 1st antibody in microplate • Washes • Block unbound sites in wells • Washes • Add antigen serum to wells • Washes • Add labeled 2nd antibody specific to antigen of interest (with substrate) • Washes • Color development • Ex: Pregnancy test

  6. Enzyme-Mediated Detection

  7. Horseradish Peroxidase EnzymeDAB Substrate DAB + H202 -------HRP-------> DAB ppt + H20

  8. AP EnzymeBCIP &NBT Substrate

  10. Process to detect HBsAg with ELISA Preparation of washing solution Add specimens (blank control, positive control, negative control)50ul and serum from patients 50ul Add enzyme: Enzyme labeled Ab 50ul(1 drop), except blank control Mixed Incubator--37℃,30mins Washing 4times Coloration: substrate A＆B.50ul--37℃,15mins Incubator

  11. Result

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