ALX 40-4C

The stably transfected cells are harvested in PBS (Ca²⁺ and Mg²⁺ free) plus 0.5 nM EDTA and washed twice with PBS. Ligand binding experiments are performed using a single concentration (0.2 nM) of ¹²⁵I-Apelin-13 in the absence or presence of increasing concentrations of unlabeled Apelin-13 or ALX 40-4C in a final volume of 100 μL of binding buffer (50 nM Hepes, pH 7.4, 1 nM CaCl₂, 5 nM MgCl₂, 0.1% bovine serum albumin) containing 5 × 10⁵ cells. Nonspecific binding is determined by the addition of 1 μM unlabeled Apelin-13. Samples are incubated for 90 min at room temperature. The incubation is terminated by separating the cells from the binding buffer by centrifugation and washing once with 500 μL of cold binding buffer. Bound ligands are determined by counting gamma emissions. At least three independent experiments are performed^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.