BIS 104 ag-ab Spring 2010

BIS 104 ag-ab Spring 2010 - Serum (cells and clotting...

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Terminology Antigen: molecule that the antibody binds to Immunogen: what an antigen is called when it is injected into an animal to induce an immune response Epitope: the specific portion of an antigen that is recognized and bound by the antibody binding site. Also know as the antigenic determinant. Process Immunization Inject antigen (+ adjuvant): IV;or IP; or IM multiple times Antigen is processed by the host animal and presented by macrophages and antigen-presenting cells (APCs) Presented to surface receptors on B-cells (Karp, p. 697 – 700) Antigen recognition by B-cells; receptor mediated activation Clonal expansion of B-cells B memory cells Plasma (Ig) secreting cells Synthesis and secretion of immunoglobulins (Ig) = antibody Multiple forms: IgG; IgM Multivalent Collect and characterize antibody Collect whole blood from immunized animal
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Unformatted text preview: Serum (cells and clotting factors removed) Fractionate out IgG from other proteins (e.g., albumin) Obtain specific antibody by affinity chromatography (fig 18.26 How does antibody react with antigen? Immunoprecipitation Agglutination Block, mask, or inhibit function How to identify and locate antigen In solution In / on cells (immunocytochemistry) In / on tissues (immunohistochemistry) On western blots (immunoblotting) Visualization of antigen – antibody interaction Label, tag antibody Radio-isotope ( 125 I; 3 H; 32 P; 35 S) (table 18.1; figs 18.20, 21) Fluorochrome (fluorescein isothyocyanate; FITC) (fig 1.12) Enzyme (horseradish peroxidase: HRP) Colloidal gold or ferritin (for immuno-electron microscopy) (fig 8.23; 9.40) Production of monoclonal antibodies (mabs): see Karp, p. 774-776 ....
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This note was uploaded on 04/16/2010 for the course MIC MIC taught by Professor Meek during the Spring '10 term at UC Davis.

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BIS 104 ag-ab Spring 2010 - Serum (cells and clotting...

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