Validating Antibodies for Quantitative Western Blot Measurements with Microwestern Array.

ABSTRACT: Fluorescence-based western blots are quantitative in principal, but require determining linear range for each antibody. Here, we use microwestern array to rapidly evaluate suitable conditions for quantitative western blotting, with up to 192 antibody/dilution/replicate combinations on a single standard size gel with a seven-point, two-fold lysate dilution series (~100-fold range). Pilot experiments demonstrate a high proportion of investigated antibodies (17/24) are suitable for quantitative use; however this sample of antibodies is not yet comprehensive across companies, molecular weights, and other important antibody properties, so the ubiquity of this property cannot yet be determined. In some cases microwestern struggled with higher molecular weight membrane proteins, so the technique

SUBMITTER: Koch RJ 

PROVIDER: S-EPMC6063895 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature