Assays to detect histoincompatbility

Question 1

Ag-Ab Interaction: The Precipitin Reaction

Answer 1

Agglutination: cross linking of epitopes on the surface of cells can induce clumping of cells
Cross linking means 1 epitope binds to more than 1 antibody–> clumping

Precipitation: cross linking epitopes on soluble antigens such as proteins can induce precipitation– soluble antigens much smaller/not whole cells – lattice like structure *Precipitation Reaction

Equivalence zone: just the right amount of antigen antibosy ratio–gives a lattice like structure

Question 2

Direct and Indirect Coomb’s Test

Answer 2

Rh- Mother with Rh+ child

Direct:
Wash fetal RBC’s covered with Maternal antibody
Add Rabit anti-human Ab
Agglutination: means that maternal ab was attached to fetal RBC

Indirect:
Take maternal serum
Add Rh+ RBC’s and wash out unbound antibody
Add Rabit anti-human Ab
Agglutination: means that maternal ab attaches to Rh+ cells

Question 3

Enzyme Linked Immunosorbent Assays (ELISAs)

Answer 3

First test for HIV

– used to detect spegic antibody.antigen in patients serum. pregnancy tests are modified ELISAs

Question 4

Western blot (immunoblot)

Answer 4

Confirmatory for HIV

ID’s antibodies

Question 5

Immunofluorescent microscopy

Answer 5

uses confocal microscope

Indirect
Binding indirectly w/ 2ndary flouresent thing to AB to ag

Direct
binding flourecent Ab to ag

Question 6

Fluorescent Activated Cell Sorting (FACS)/Flow Cytometry

Answer 6

Machine that determines frequency of a given cell type in a heterogenous mixture

done for CDCs–> WBC counting
helps w/ AIDs diagnosis

Question 7

Monoclonal Antibody Production

Answer 7

making antibodies to specific receptors for targeted immunotherapy