A Path to Protein Quantitation: PBMCs to Western Blot

Abstract

Proteins are biopolymers made up of one or more long chains of amino acid residues. These proteins mediate thousands of metabolic pathways that determine how an organism functions. When proteins are thoroughly investigated, they can provide information about health and disease. A variety of methods have been developed to quantify both complex protein mixtures for total protein content as well as a single type of protein. This manuscript covers the methodologies standardized in our lab based on previously published protocols, beginning with the isolation of peripheral blood mononuclear cells (PBMCs) from human blood samples, cell viability testing, cell counting, total protein extraction methods, and protein quantification and estimation techniques, which enable a researcher to find the protocol from PBMCs to Western blot in a single manuscript.