Immunology lecture 03
Terms
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- What is ELISA?
- A solid phase immunoassay in which protein is absorbed to plastic and antibody-antigen interactions are qualitatively or quantitatively measured, based on light absorbance
- What is Radioimmunoassay (RIA)
- Essentially an ELISA that uses radioactivity in place of the enzyme to allow detection of the bound antibody
- What is RIA useful for?
- RIA is highly sensitive, so it is particullarly useful for the quantification of low-abundance substances
- What is FACS (flow cytometry)
- Most frequently used for the analysis of leukocytes, although any cell type that can be prepared as a suspension can be analyzed
- What makes FACS (flow cytometry) powerful?
- Ability to quantitatively analyze large numbers of cells, and to accurately identify complex or small populations. Also possible to separate specific populations of cells, based on specific phenotypes
- What is immunofluorescence?
- Similar to FACS, but fluorescence detection is by microscopy. Only a small number of cells can by analyzed
- When would immunofluorescence be used?
- To analyze or identify subcellular structures, to identify specific populations of cells, or to analyze a biopsy in which the available cell population is very small
- What does Western blot do?
- Fractionates proteins based on size
- What is the Coombs test used for?
- Testing for anti-Rh antibodies and to diagnose immune-mediated hemolytic anemia. Specifically detects antibodies against red blood cells
- What is the direct Coombs test
- Red blood cells are recovered from a patient via blood draw, and mixed with the reagent. Positive test is agglutination
- What is the indirect Coombs test
- Red blood cells are mixed with the patient and then with the Coombs reagent. Commonly used for pregancy
- Name three sources of antibodies
-
Human Serum (used to measure a patient's antibody response)
Polyclonal antibodies (used as detection reagents)
Monoclonal antibodies (hybridomas) - What is a proliferation assay used for
- To measure the ability of B cells or T cells to proliferate in response to mitogenic stimuli
- What is a functional assay used for
- To measure specific effector functions of leukocytes
- Name two nucleic acid-based assays
- Southern blot and Microarrays