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10 Cards in this Set
- Front
- Back
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What do you use RIA and ELISA together to measure?
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quantify molecules in serum
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How do you conduct the RIA and ELISA?
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1) Add anti-A antibody covalently linked to an enzyme
2) Wash away unbound antibody 3) Enzyme will make a colored product when you add a substrate 5) Measure absorbance of light of colored product |
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What is the general principle behind the ELIZA?
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plate bound antibodies capture molecules which are in turn detected by labelled antibodies recognizing a different epitope
values are compared to a standard curve |
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What is the principle behind agglutination and hemagluttination?
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antibodies directed against an antigen on the surface of a particule will cause clumping. measure the dilution of serum required to cause clumping is called the titre
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↑ titre: ____ concentration of antibody in serum
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↑ titre: ↑ concentration of antibody in serum
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What happens in the Combs test?
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hemagglutination for prescence of Rh antibody
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How do you produce monoclonal antibodies?
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1) fuse myeloid cells with immortal myeloma cells with a enzyme
2) plate them with the substrate 3) only those cells with enzyme will survive so you know they are the hybridoma cells 4)select hybridoma cells that make antibodies to what you are looking for 5) clone that hybridoma |
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What do you use to detect antigens in situ?
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immunoflourescence microscopy
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How do you do immunoflouresence microscopy?
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antibodies labelled with flourochorme are put into serum then light is transmitted
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What are the general principles of flow cytometry?
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1) flourochrome labelled antibodies are used to detect antigens on/in individual cells in a mixture
2) cells expressing antigens are detected by their ability to emit a specific wavelength of light 3) detectors can also measure scatter of laser as a measurement of other characteristics of the cell 4) cells can be separated using charged plates |
