Assays Include Those Using Biochemical Reagents Like Receptors or Antibodies
Isolate/prepare a receptor suspension,
or a serum binding protein, or an antibody
Isolate/prepare a small amount of purified
Need to label part of it so it can be visualized
Need pure hormone for standards or references,
In each of a series of tubes or wells place
the same small amount of label and binder - the binder will be the limiting
reagent in this assay
In part of this series of tubes add increasing
amounts of unlabeled hormone, one amount in each tube or well starting
at zero additional.
In the rest of this series of tubes add
a measured volume of the unknowns, one unknown per tube.
Incubate for a period of hours
Separate the hormone bound to the binder
from the remaining free hormone by some simple method.
Quantify the amount of label that is bound
to the binder; it will be inversely proportional to the amount of unlabeled
hormone present since the binder is the limiting reagent - this is a molecular
game of musical chairs.