Aortic tissue segments were opened longitudinally and incubated with PBS-T containing 10% donkey serum. Tissue segments were then rinsed 3 times with PBS-T and incubated overnight at 4 °C with anti-mouse VCAM-1 polyclonal antibodies (Santa Cruz Biotechnology Inc; 1:100) diluted in PBS-T. Anti-donkey secondary antibody labeled with Alexa Fluor fluorophore molecules (Molecular Probes Inc.) was then applied for 45 min at room temperature. The segments were mounted on slides, covered with Vectorshield mounting medium (Vector Laboratories Inc) and observed with a FV1000 confocal system (Olympus). At least 4 animals were used. Five pictures of each field were captured at different focal lengths and VCAM-1 positive cells per 1000 endothelial cells were counted.
Alexa fluor fluorophore molecules
Manufactured by Thermo Fisher Scientific
Alexa Fluor fluorophore molecules are a series of fluorescent dyes developed by Thermo Fisher Scientific. These dyes are designed to provide bright, photostable fluorescence for a variety of applications in biological research. The core function of Alexa Fluor fluorophores is to serve as fluorescent labels that can be conjugated to various biomolecules, such as proteins, nucleic acids, or other molecules of interest, to facilitate their detection and visualization in various experimental techniques.
Automatically generated - may contain errors
2 protocols using alexa fluor fluorophore molecules
1
Immunohistochemical Analysis of Aortic VCAM-1
2
Immunohistochemical Analysis of Aortic VCAM-1
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Aortic tissue segments were opened longitudinally and incubated with PBS-T containing 10% donkey serum. Tissue segments were then rinsed 3 times with PBS-T and incubated overnight at 4 °C with anti-mouse VCAM-1 polyclonal antibodies (Santa Cruz Biotechnology Inc; 1:100) diluted in PBS-T. Anti-donkey secondary antibody labeled with Alexa Fluor fluorophore molecules (Molecular Probes Inc.) was then applied for 45 min at room temperature. The segments were mounted on slides, covered with Vectorshield mounting medium (Vector Laboratories Inc) and observed with a FV1000 confocal system (Olympus). At least 4 animals were used. Five pictures of each field were captured at different focal lengths and VCAM-1 positive cells per 1000 endothelial cells were counted.
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