User manual

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10.3.1.1 Will the Antagonist Rapidly Unbind?
For a typical antagonist/agonist experiment, whether or not the antagonist rapidly unbinds has a large
effect on the number of perfusion solutions (and valves) needed to get a good dose-response curve.
If the antagonist remains bound during the application of a solution containing agonist only, then
antagonist does not need to be added to the agonist solution to get an accurate dose-response curve.
Therefore, you can have separate solutions (and valves) for each agonist solution as well as each
antagonist solution.
For example, one eight valve controller is sufficient to do a dose-response curve for 4 agonist
concentrations and 4 antagonist concentrations (one of which could be no antagonist, ie ACSF) (Table
10.3.1.1.1).
Antagonist Agonist
Ch 1 1
Ch 2 2
Ch 3 3
Ch 4 4
Ch 5 1
Ch 6 2
Ch 7 3
Ch 8 4
Table 10.3.1.1.1. If the antagonist does not rapidly unbind, eight solutions are sufficient to produce a
dose-response curve for 4 agonist concentrations and 4 antagonist concentrations (one of which could be
no antagonist, ie ACSF). This can be done with one eight channel controller controlling eight valves.
If the antagonist does not remain bound during the application of the agonist-only solution, then the
antagonist has to be added to the agonist solution to get an accurate dose-response curve. Therefore,
many more agonist + antagonist solutions (and valves) are required to do a dose-response curve.
For example, to do a dose response curve for 4 agonists and only 3 antagonists, 15 solutions and 15
valves (two eight valve controllers) is required (Table 10.3.1.1.2, see also Fig. 10.3.1.2.4)