CSB351Y1 Lecture Notes - Lecture 5: Microtiter Plate, Hypersensitive Response, Chromogenic
18 Oct 2016
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Infection unit (iu): smallest amount of virus capable of producing a reaction. Multiplicity or infection (moi): the ratio of input virus particles/host ratio. Hypersensitive response to tmv: either a systemic or a local lesion response to infected parts of leaf. Make a series of dilutions and rub an aliquot onto a leaf; count lesion number. Each infectious particle produces a circular zone of infected cells called a plaque. Similar to local lesion assay, except in animals and bacteria. Add sample of known ag (antigen) of known concentration to a well of a microtiter plate. Then add bovine serum albumin (bsa) or casein for blocking; used to block other irrelevant proteins from binding to the wells. Specific ag of interest is applied to all plate wells. Blood sample is added (ag-ab complexes are formed) Secondary antibodies will bind to any ag-ab complexes. Apply a substrate which is converted by the enzyme to elicit a chromogenic or fluorogenic or electrochemical signal.
