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.

ABSTRACT:

Background

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 CB₁ receptor in particular show a progressive tendency to aggregate and precipitate in aqueous solution outside of their biological context due to the low solubility that the hydrophobic nature imprinted by their seven transmembrane domains. This renders full-length recombinant GPCRs useless for analytical purposes, a problem that can be overcome by engineering soluble recombinant fragments of the receptor containing the antigen.

Results

Here we generated highly soluble and stable recombinan

SUBMITTER: Saumell-Esnaola M 

PROVIDER: S-EPMC9479267 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature