Immunologic Tests--Diebel Flashcards
HAT media
Hypoxanthine: converted by hypoxanthine phosphoribyl transferse (HGPRT)
–> specific for fused cells
–> rescue for fused cells
Aminopterin: inhibits DHFR, inhibits nucleotide synthesis
Thymidine: rescue
Hypoxanthine
hypoxanthine phosphoribyl transferase converts to GTP/ATP
–> only in fused cells, not myeloma cells
–> differential sensitivity
–> rescue
–> bypasses DHFR
Aminopterin
inhibits DHFR, inhibits nucleotide synthesis
Thymidine
rescue
Immunoprecipitation assays
visualize precipitation of antibody/antigen complexes
Direct agglutination assays
visualize clumped cells w/ Fab bound
Indirect agglutination assays
visualize latex beads
–> latex beads bound to Fc region
–> when antibody binds Fab, Fc regions clump together
–> we actually see the latex beads grouping together
Neutralization assays are based on the principle of…
cell survival
*antibodies against toxin are added
+ test = cell survives
shows that toxin was present
- test = cell dies
try again
Complement fixation assays
testing if complement is available
- test = hemolysis
*complement is available
+ test = no hemolysis
*complement is not available
Direct vs indirect immunofluorescence
direct = tagged antibody is specific for antigen
indirect = tagged antigen is generic
Direct ELISA
looking for antigen
antibody is in plate
serum added (with antigen?)
enzyme-linked antibody is added
enzyme substrate is added
+ = color change
Indirect ELISA
looking for antibody
antigen present in well
serum (with antibody?) added to well
enzyme-linked antibody added
substrate added
positive test = change
Competative ELISA
look at your chart, this one is difficult
ELISPOT looks for
cytokines
