The Bicinchoninic Acid (BCA) Assay for Protein Quantitation John M. Walker 1. Introduction The bicinchoninic acid (BCA) assay, first described by Smith et al. (1) is similar to the Lowry assay, since it also depends on the conversion of Cu2+ to Cu+ under alkaline conditions (see Chapter...
The Bicinchoninic Acid (BCA) Assay For Protein Quantitation John M. Walker 1. Introduction The bicinchoninic acid (BCA) assay, first described by Smith et al. (1) is similar to the Lowry assay, since it also depends on the conversion of Cu2+ to Cu+ under alkaline conditions (see Chapter...
(1) is similar to the Lowry assay, since it also depends on the conversion of Cu2+to Cu+under alkaline conditions (seedoi:10.1385/0-89603-268-X:5John M. WalkerMethods Mol BiolWalker, J.M., 1996. The bicinchoninic acid (BCA) assay for protein quantification. In: Walker, J.M. (Ed...
The bicinchoninic acid assay (BCA assay), Aminolink Plus coupling resin and spin columns were purchased from Thermo Scientific (Rockford, Ill). RBC preparation RBCs were prepared for experiments under different oxygen saturations using previously established protocols. Fresh blood was collected from ...
Protein concentrations were quantified with a bicinchoninic acid (BCA) protein assay kit from Thermo Fisher Scientific, Inc. (Waltham, Massachusetts, United States). For protein separation, 2 μg of each sample underwent 12% sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE). ...
The concentration of total protein was determined by using the Bicinchoninic Acid Protein Assay Kit (BCA1, B9643, Sigma-Aldrich) according to the manufacturer’s instructions. Transmission electron microscopy The mouse peritoneal cells were pelleted by centrifugation at 3000 r.p.m. and then ...
a Bicinchoninic acid (BCA) in vitro enzyme assay. The purple copper bicinchoninate complex indicating GlgX-mediated generation of reducing ends in glycogen was quantified by photometric measurements at 562 nm. A562 values were converted into reducing ends [µM glucose equivalents] based on d-...
Protein content of lysates was determined using the bicinchoninic acid assay (BCA™, Pierce, Rockford, IL, USA). Samples were separated by SDS-PAGE with precast gels (4-15%, Bio-Rad) and subsequently the proteins were transferred to nitrocellulose membrane (Hybond-ECL, Amersham) with a ...
The protein concentrations of the supernatant were determined using the bicinchoninic acid (BCA) Protein Assay Kit (Thermofisher Scientific) and stored at −80C. An equal number of proteins (30 μg/well) were separated (10 % SDS-PAGE) and transferred to a polyvinylidene fluoride (PVDF) ...
Protein concentration was determined using the bicinchoninic acid (BCA) assay (BioRad). To measure ceramide synthase activity, a fluorescent assay was conducted as described by Kim et al. (Kim et al., 2012a) with minor modifications. Briefly, 50 µg homogenate protein was incubated in ...