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chapt 6


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antibody affinity
strength of total noncovalent interactions between SINGLE Ag binding site on Ab and SINGLE epitope
preferred parameter for affinity description
what scatchard plot reveals
Ka (slope)

Valency of antibody
a better description of the binding capability of an antibody; takes into account both affinity and valency.
what type of autoimmune reaction results from Streptococus pyogenes?
these express M proteins which cross react with myocardial proteins; heart is damaged.
Are precipitation reactions high or low sensitivity?
2 requirements of precipitation reaction
bivalent antigen, bivalent antibody
according to her ouchterlony is radial or double?
steps in radioimmunoassay
1. immobilize antibody on plate
2. add radiolabeled antigen to saturate binding sites.
3. add natural sample to compete for binding sites.
4. precipitate radio-antigen-antibody complexes to measure radioactivity; the amount it decreased from originally is how much sample is present.
indirect elisa tests for:
ANTIBODY in the serum!! immolize antigen on plate, add sample for testing; add secondary antibody w/ enzyme, then substrate for cleaving. measure.
sandwich elisa tests for:
ANTIGEN!! mobilize antibody on plate; add sample, it binds; add antibody to the other epitopes on the Ag, containing the enzyme to cleave. that's why the antigen has to be multiepitoped
competitive elisa tests for:
ANTIGEN; mix sample with antibody first!

then add ag-ab MIXTURE (including free antibody) to immobilized antigen on plate.

if you have less antigen in sample, more antibody will be free to bind antigen on plate. add 2Antibody with enzyme, it binds to antibody sticking to well's antigens. inversely prptnl
how do you measure size and quantity of protein?
western blot
how do you purify protein?

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