Quantipro bca assay kit

Sample were added 100 μl of lysates on ice for 30 min. The cell and exosomes lysates were clarified by centrifugation at 12,000 rpm for 15 min, and the supernatants were collected. The protein concentration was measured with the QuantiPro BCA Assay Kit (KeyGen Biotech Co., Ltd., Shanghai, China). The protein concentration of each sample was measured with the QuantiPro BCA Assay Kit (KeyGen Biotech, Shanghai, China). 20 μg protein was applied to Western Blotting. The membranes were incubated overnight at 4°C with specific anti-CD63 (diluted 1: 200; Abcam, United States), anti-CD81 (diluted 1: 500; Abcam, United States), anti-CD40 (diluted 1: 1000; Bioss, China), anti-ALIX (diluted 1: 1000; Abbexa, United Kingdom), anti-RUNX2 (diluted 1: 500, SAB, United States), anti-BMP-2 (diluted 1: 500, Bioworld, United States), anti-OPN (diluted 1: 1000, Proteintech, United States), and anti-GAPDH (diluted 1: 5000, Bioworld, United States). Incubation with the secondary antibody (diluted 1: 500, ABclonal, China) lasted 1 h. The ECL luminescent solution was configured to collect the blotting results with a BIO-RAD gel imaging system, and the results were analyzed with Image Lab software.

The protein concentration of each sample was measured using a QuantiPro BCA Assay Kit (KeyGen Biotech, Shanghai, China). The membranes were then incubated overnight at 4 ºC with specific anti-CD63 (diluted 1: 200; Abcam, Cambridge, MA, USA), anti-CD81 (diluted 1: 500; Abcam), anti-CD40 (diluted 1: 1000; Bioss Antibodies, Woburn, MA, USA), anti-ALIX (diluted 1: 1000; Abbexa Ltd., Cambridge, UK), anti-RUNX2 (diluted 1: 500, SAB, USA), anti-BMP-2 (diluted 1: 500, Bioworld Technology, St Louis Park, MN, USA USA), anti-ALP (diluted 1: 1000, Abcam), anti-GJA1 (diluted 1: 1000, Bioworld), anti-AP3B1 (diluted 1: 300, Proteintech, Rosemont, IL, USA) and anti-GAPDH (diluted 1: 5000, Bioworld). Incubation with the secondary antibody (diluted 1:500, ABclonal, Woburn, MA, USA) lasted 1 h. The ECL luminescent solution was configured to collect the blotting results with a Bio-Rad gel imaging system (Bio-Rad, Hercules, CA, USA), and the results were analyzed using Image Lab software.