BIOL 620 1st Edition Lecture 11 Outline of Last Lecture I. Serology Outline of Current Lecture II.Immuno diffusion assayIII. Double immuno diffusion assayCurrent LectureImmunology: Lecture 12- Immuno diffusion assay:This refers to detecting Ab.Ag reaction with an extensive lattice formation.In an agar plate, 2 wells are formed one with Ab and alpha BSA (anti serum) and other with BSA and Ag. After an overnight wait, a precipitation line forms in the center. This is due to diffusion in all directions asthe same concentrations of Ag and Ab are used. Using a graph where the Ag line crosses the Ab line is the equivalence point. Equivalence point is the right ratio of the Ag.Ab to form precipitate.In another agar plate, the concentration of Ag is half the concentration of Ab. Here, the precipitation lineis closer to Ag well than the Ab well.When size is altered whereby, Ag1> Ag2, one well has alpha Ag 1 and alpha Ag2. Another well has Ag1 and Ag2. There are 2 precipitation lines, one for Ag1.Ab1 and other for Ag2.Ab2. Since diffusion depends on size, large size diffuses slower than the smaller one.These notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.Double immuno diffusion assay: Aka Ouchterlong assay:This is used to analyze epitope structures.Ask:1. Does Ab recognize epitopes on Ag? 2. Will it form precipitate or extensive lattice?3. How do set of epitopes on neighboring wells compare to each other?Manipulate Ab by absorption by removal of specificity: Absorbed on BSAAlpha (BSA.DNP) ---------------------- Alpha DNP only. No precipitate
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