Neuroproteomics by Firas H. Kobeissy & Stanley M. Stevens Jr

Author:Firas H. Kobeissy & Stanley M. Stevens, Jr.
Language: eng
Format: epub
Publisher: Springer New York, New York, NY

Saturation Binding Assay (to determine the concentration of agonist binding sites)

The following procedure is for a centrifugation-based binding assay with increasing concentrations (2–100 nM) of a high affinity [3H]ligand. Each condition will be done in duplicate with a final volume of 100 μL and a final protein concentration of 0.25 mg/mL per condition.1.Prepare stock solution of [3H]ligand (200 nM).

2.Prepare 2500 μL of 0.5 mg/mL pLGIC preparation. A minimum quantity of receptor per condition is used to ensure saturation of receptor sites within the [3H]ligand concentration used.

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