Dopamine Receptor Binding and Quantitative Autoradiographic Study - Drugs of Abuse: Neurological Reviews and Protocols - page 300

Radioligand concentrations

A B C

Mean Working Final

dpm conc. ( M ) conc. (n M )

1 1 254585 9.80E-09 1.96

1 2 156335 6.02E-09 1.20

1 3 1 92810 3.57E-09 0.71

1 4 1 57685 2.22E-09 0.44

1 5 1 34080 1.31E-09 0.26

1 6 1 21235 8.18E-10 0.16

1 7 1 12935 4.98E-10 0.10

1 8 11 7520 2.90E-10 0.06

1 9 11 4830 1.86E-10 0.04

10 11 2800 1.08E-10 0.02

Binding

Protein conc.: 245 µ g/tube

A A D E F G

Radioligand Total Nonspecifi c Specifi c Specifi c Specifi c

conc. (n M ) (dpm) (dpm) (dpm) (mol) (fmol/mg of protein) B/T

1.96 4317 3060 1257 4.84E-15 19.75 10.08

1.20 3100 1824 1276 4.91E-15 20.05 16.66

0.71 2173 1012 1161 4.47E-15 18.24 25.53

0.44 1635 1 615 1020 3.93E-15 16.03 36.09

0.26 1207 1 397 1 810 3.12E-15 12.73 48.51

0.16 1 805 1 241 1 564 2.17E-15 1 8.86 54.20

0.10 1 607 1 158 1 449 1.73E-15 1 7.06 70.84

0.06 1 434 1 104 1 330 1.27E-15 1 5.19 89.56

0.04 1 296 11 82 1 214 8.24E-16 1 3.36 90.42

0.02 1 190 11 58 1 132 5.08E-16 1 2.07 96.21

Data are those resulting from saturation analysis using [ 3 H]PD 128907 (specifi c activity =
117 Ci/mmol) in rat ventral striatal membranes (protein concentration 245 µg/tube). Counts
from the working solutions of radioligand and total and nonspecifi c binding are obtained from
the scintillation counter and represent the mean of duplicate determinations. All other values are
calculated from these data. Graphical representation of this data is shown in Fig. 2 .
A: Average of duplicates from scintillation counter expressed in dpm.
B: Working conc. ( M ) =

dpm

————————————————
2.22e12 dpm/Ci *117 Ci/mmol * 0.1 mL
C: Final conc. (n M ) = Working conc. (M) * e9l

—————————-
5
D: Specifi c binding (dpm) = Total binding (dpm) - nonspecifi c binding (dpm).
E: Specifi c binding (mol) =

Specifi c binding (dpm)

———————————————————
2.22e12 dpm/Ci *117Ci/mmol*1e-3 mmol/mol
F: Specifi c binding (fmol/mg of protein) = Specifi c binding (mol) * 1e15 fmol/mol

————————————————

0.245 mg of protein/tube

G: B/ T = Specifi c binding (fmol/mg of protein)

———————————————

Radioligand concentration (n M )