Mouse monoclonal antibody against cd31

An immunofluorescence analysis was carried out to identify blood vessel formation. Blood vessel staining was performed with a mouse monoclonal antibody against CD31 (1:100; Abcam, Cambridge, MA, USA) and a rabbit polyclonal antibody against alpha smooth muscle actin (α-SMA; 1:200; Abcam), with the following secondary antibodies: anti-rabbit Alexa Fluor^® 594 (A21207; 1/1,000; Thermo Fisher Scientific) and biotin–streptavidin Alexa Fluor^® 488 (BA-2001, 1:200; Thermo Fisher Scientific – 532354, 1:150, Thermo Fisher Scientific). For confocal imaging, an LSM710 Meta confocal microscope (Carl Zeiss, Feldbach, Switzerland) was used. α-SMA and CD31-positive structures were identified and photographed. Arterioles and capillaries bordering the infarct at the mid-infarct level were identified by counting the α-SMA- and CD31-positive structures, subsequently computed using a fluorescence microscope in a blinded manner from eight randomly selected images as the mean within 1-mm² areas. The maturation index was calculated based on the number of α-SMA-positive vessels relative to the total number of vessels.28 (link) All data analyses were performed in a blinded manner. All measurements were performed using Image J software.