logging in or signing up Immunological Techniques deepthi12 Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINT lite Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: Embed: Flash iPad Copy Does not support media & animations WordPress Embed Customize Embed URL: Copy Thumbnail: Copy The presentation is successfully added In Your Favorites. Views: 9334 Category: Education License: All Rights Reserved Like it (5) Dislike it (1) Added: June 22, 2009 This Presentation is Public Favorites: 7 Presentation Description about different kind of immunological techniques Comments Posting comment... Premium member Presentation Transcript Slide 1: Antigen-Antibody Interactions: Principles & Applications --by K.R.Deepthi - A bimolecular association involving various noncovalent interactions Is similar to an enzyme-substrate interactions, but not lead to an irreversible chemical alteration Slide 2: Nature of Ag/Ab Reactions Slide 3: Strength of Antigen-Antibody Interactions Precipitation Reactions Agglutination Reactions Immunodiffusion Radioimmunoassay Enzyme-Linked Immunosorbent Assay Western Blotting/immunoblotting Immunofluorescence Immunohistocompatibility Localization of cells in tissue immunoblotting. contents: Structure of an antibody : Structure of an antibody Slide 5: Affinity = ? attractive and repulsive forces Affinity Strength of the reaction between a single antigenic determinant and a single Ab combining site Calculation of Affinity : Calculation of Affinity Ag + Ab ? Ag-Ab Applying the Law of Mass Action: Slide 7: Avidity The overall strength of binding between an Ag with many determinants and multivalent Abs Specificity : Specificity The ability of an individual antibody combining site to react with only one antigenic determinant. Slide 9: Cross Reactivity The ability of an individual Ab combining site to react with more than one antigenic determinant. The ability of a population of Ab molecules to react with more than one Ag Slide 10: Factors Affecting Measurement of Ag/Ab Reactions Affinity Avidity Ag:Ab ratio Physical form of Ag Slide 11: Precipitation Reactions Radial Immunodiffusion (Mancini) : Radial Immunodiffusion (Mancini) Interpretation Diameter of ring is proportional to the concentration Quantitative Ig levels Method Ab in gel Ag in a well Slide 14: Precipitation Reactions(immunoelectrophorosis) Countercurrent electrophoresis : Countercurrent electrophoresis Method Ag and Ab migrate toward each other by electrophoresis Used only when Ag and Ab have opposite charges Qualitative Rapid Slide 16: Agglutination/Hemagglutination Definition - tests that have as their endpoint the agglutination of a particulate antigen Agglutinin/hemagglutinin Slide 17: FIGURE 6-7 Demonstration of hemagglutination using Ab against sheep red blood cells (SRBCs). Agglutination Reactions visible clumping by interaction between Ab & a particulate antigen such as RBC, latex beads. -routinely performed to type RBCs for blood transfusion. Slide 18: RIA Slide 19: From these data, a standard binding curve, like the one shown in red, can be drawn. RIA RIA : RIA Radioimmunoassay is widely-used because of its great sensitivity. Using antibodies of high affinity, it is possible to detect a few picograms (10-12 g) of antigen in the tube. The greater the specificity of the antiserum, the greater the specificity of the assay Slide 21: ELISA Slide 22: FIGURE 6-12 Western blotting : separates the components according to their molecular weight. : the proteins in the gel are transferred to the sheet of nitrocellulose or nylon by the passage of an electric current. : probed with Ab & then radiolabeled or enzyme-linked 2nd Ab. : a position is visualized by means of an ELISA reaction. Slide 23: Immunofluorescence mIgM-producing B cells indirectly stained with rhodamine-conjurated secondary Ab under a fluorescence microscope. FIGURE 6-14 Localization of cells in tissue immunoblotting : Localization of cells in tissue immunoblotting Nonsymbiotic hemoglobins (ns-Hbs) previously have been found in monocots and dicots. however, very little is known about the tissue and cell type localization as well as the physiological function(s) of these oxygen-binding proteins. The immunodetection and immunolocalization of ns-Hbs in rice (Oryza sativa L.) by Western blotting and in situ confocal laser scanning techniques. Ns-Hbs were detected in soluble extracts of different tissues from the developing rice seedling by immunoblotting. Levels of ns-Hbs increased in the germinating seed for the first six days following imbibition and remained relatively constant thereafter. Immunohistocompatibility : It is similar to grafting Immunohistocompatibility You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.