. Author manuscript; available in PMC: 2016 Dec 8.

Published in final edited form as: Curr Protoc Pharmacol. 2015 Dec 8;71:1.34.1–1.3421. doi: 10.1002/0471141755.ph0134s71

Table 1.

Sample dilution scheme of [³H]-(+)-pentazocine for saturation-binding experiments

Dilution #	Final [³H]-(+)-PTZ concentration¹ (nM)	10X stock [³H]-(+)-PTZ concentrations (nM)	Dilution factor	Dilution volumes
1	300 nM	3000 nM	10	102 μL stock + 898 μL of Assay Buffer
2	100 nM	1000 nM	28	34 μL of stock + 966 μL of Assay Buffer
3	30 nM	300 nM	10	100 μL of dilution #1 + 900 μL of Assay Buffer
4	10 nM	100 nM	10	100 μL of dilution #2 + 900 μL of Assay Buffer
5	3 nM	30 nM	10	100 μL of dilution #3 + 900 μL of Assay Buffer
6	1 nM	10 nM	10	100 μL of dilution #4 + 900 μL of Assay Buffer
7	0.3 nM	3 nM	10	100 μL of dilution #5 + 900 μL of Assay Buffer

Final radiolgiand concentration is the concentrations of [³H]-(+)-pentazocine in the binding assay.

² The stock concentration of [³H]-(+)-pentazocine is supplied at 34 Ci/mmol (e.g., Perkin Elmer). The calculated concentrations of this stock [³H]-(+)-petnazocine is counted and determined be approximately 29.4 μM (1/34 Ci/mmol × 1000) which is the concentrations used to prepare the 10X stocks radioligand.