Design and validation of recombinant protein standards for quantitative Western blot analysis of cannabinoid CB1 receptor density in cell membranes: an alternative to radioligand binding methods

• Published in: Microbial cell factories Vol. 21; no. 1; pp. 1 - 192
• Main Authors: Saumell-Esnaola, Miquel, Elejaga-Jimeno, Ainhoa, Echeazarra, Leyre, Borrega-Román, Leire, Barrondo, Sergio, López de Jesús, Maider, González-Burguera, Imanol, Gómez-Caballero, Alberto, Goicolea, María Aranzazu, Sallés, Joan, García del Caéo, Gontzal
• Format: Journal Article
• Language: English
• Published: London BioMed Central Ltd 15.09.2022; BioMed Central; BMC
• Subjects: Antibodies; Antigens; Aqueous solutions; Binding; Binding sites; Biological properties; Biological samples; Cannabinoid CB1 receptor antibodies; Cannabinoid CB1 receptors; Cannabinoids; Carboxy-terminal tail; Cell membranes; Cell receptors; Chemical properties; Cloning; Context; Cost analysis; Density; Design; G protein-coupled receptors; GPCR expression analysis; Homology; Hydrophobicity; Ligands; Medical research; Membrane proteins; Membranes; Methods; Peptides; Pharmaceutical industry; Physiological aspects; Plasmids; Production processes; Protein engineering; Protein expression; Proteins; Quantitative analysis; Quantitative Western blot; Radioactivity; Radioligand saturation binding; Receptor density; Receptors; Recombinant proteins; Residues; Saturation; Soluble recombinant protein standards; Synaptosomes; Transmembrane domains; Western immunoblotting; Spain

Replacement of radioligand binding assays with antibody-antigen interaction-based approaches for quantitative analysis of G protein-coupled receptor (GPCR) levels requires the use of purified protein standards containing the antigen. GPCRs in general and cannabinoid CB.sub.1 receptor in particular s...