Quick start bradford protein microplate standard assay

Protein concentrations in extracellular protein extract and cell lysate were estimated by Quick Start^TM Bradford protein microplate standard assay (Bio-Rad) as per manufacturer instructions.

Protein concentrations in the EVs and WCP samples were determined by Quick Start^TM Bradford protein microplate standard assay (Bio-Rad, USA). For protein separation, the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using the mini-PROTEAN II cell electrophoresis system (Bio-Rad, USA). The proteins were denatured in 2× loading buffer at 100°C for 5 minutes, followed by centrifugation at 5000 ×g for 5 minutes. 20 μl of proteins loaded in each well of the gel were separated on 12% SDS-PAGE at a constant 120 V. After the run was completed, protein bands were detected using silver stain. Gel images were visualized in G: Box Imaging System (Syngene, India). Protein banding patterns and molecular weights of the bands were determined using GeneSys tools software.