Goat antirabbit mouse hrp linked secondary antibody

All proteins from cells or myocardial tissue were extracted and quanti ed. Micro BCA TM Protein Assay Kits (Thermo Fisher Scienti c, Waltham, MA) were used to quantify protein levels. Equivalent amounts of protein was electrophoresed through 12% SDS-PAGE (stacking gel, 70V; separating gel, 110V) and transferred to nitrocellulose membranes (200 mA, 50 minutes). Blots were incubated for 1 hour at room temperature with the indicated antibodies (CD63: ab108950, 1:1000, Abcam, CA, USA; TSG101: ab125011, 1:5000, Abcam; Cleaved-Caspase-3: ab2302, 1:500, Abcam; Bcl-2: ab 196495, 1:1000, Abcam; Bad: ab32445, 1:5000, Abcam; Bax: ab32503, 1:5000, Abcam), and then incubated with goat-antirabbit/mouse HRP-linked secondary antibody (Abcam, Cambridge, MA, USA). Chemiluminescence substrate (Pierce Chemical, Rockford, IL, USA) was used to visualize protein signals. The intensity of the protein bands was analyzed by Image J software (NIH, USA).

All proteins from cells or myocardial tissue were extracted and quanti ed. Micro BCA TM Protein Assay Kits (Thermo Fisher Scienti c, Waltham, MA) were used to quantify protein levels. Equivalent amounts of protein was electrophoresed through 12% SDS-PAGE (stacking gel, 70V; separating gel, 110V) and transferred to nitrocellulose membranes (200 mA, 50 minutes). Blots were incubated for 1 hour at room temperature with the indicated antibodies (CD63: ab108950, 1:1000, Abcam, CA, USA; TSG101: ab125011, 1:5000, Abcam; Cleaved-Caspase-3: ab2302, 1:500, Abcam; Bcl-2: ab 196495, 1:1000, Abcam; Bad: ab32445, 1:5000, Abcam; Bax: ab32503, 1:5000, Abcam), and then incubated with goat-antirabbit/mouse HRP-linked secondary antibody (Abcam, Cambridge, MA, USA). Chemiluminescence substrate (Pierce Chemical, Rockford, IL, USA) was used to visualize protein signals. The intensity of the protein bands was analyzed by Image J software (NIH, USA).