As described above, the cavernosal tissue was obtained immediately after we performed the cavernosometery. The same quantity of protein extracts (20–50 µg) were separated in sodium dodecyl sulfate polyacrylamide gels, which were then transferred to nitrocellulose membranes. An overnight incubation was performed with primary antibodies, including anti-neuronal nitric oxide synthase (1:2,000; Cell Signaling Technology, Boston, MA, USA), anti-endothelial nitric oxide synthase (eNOS) (1:3,000; BD Biosciences, San Jose, CA, USA), anti-phospho-eNOS (Ser1177, 1:1,000; Cell Signaling Technology), anti-myosin phosphatase target subunit 1 (MYPT1) (1:2,000; Cell Signaling Technology), anti-phospho-MYPT1 (Thr696, 1:1,000; Millipore, Charlottesville, VA, USA), and platelet endothelial cell adhesion molecule-1 (PECAM-1) (1:1,000; Santa Cruz Biotechnology) [3 (link)4 (link)6 (link)7 (link)13 (link)16 (link)]. The bands were visualized using the electrochemiluminescence Western blotting development system. To adjust for any loading differences, the membranes were re-probed with an antibody against monoclonal anti-actin antibodies. The results were then quantified by means of densitometry using Image J analysis software (National Institute of Health).
Anti endothelial nitric oxide synthase
Manufactured by BD
Sourced in United States
Anti-endothelial nitric oxide synthase (eNOS) is a laboratory equipment product used for the detection and quantification of the enzyme endothelial nitric oxide synthase. eNOS is responsible for the production of nitric oxide, which plays a crucial role in various physiological processes. This product allows researchers to measure eNOS levels and activity in biological samples.
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Lab products found in correlation
Anti phospho enos ser1177, by Cell Signaling Technology (1 mentions)
Anti phospho mypt1 thr696, by Merck Group (1 mentions)
Anti neuronal nitric oxide synthase, by Cell Signaling Technology (1 mentions)
Pecam 1, by Santa Cruz Biotechnology (1 mentions)
Magnetic column, by Miltenyi Biotec (1 mentions)
Fibronectin coated chamber slides, by BD (1 mentions)
Streptavidin conjugated magnetic beads, by Miltenyi Biotec (1 mentions)
Anti von willebrand factor, by Santa Cruz Biotechnology (1 mentions)
Dii ac ldl, by Thermo Fisher Scientific (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
2 protocols using anti endothelial nitric oxide synthase
1
Cavernosal Protein Expression Analysis
2
Magnetic Separation of CD45+ Cells
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Cells derived from high density culture at day 15 were stained with a biotinylated anti-rat-CD45 monoclonal antibody (BD Biosciences) for 30 min on ice, washed, and then incubated with streptavidin-conjugated magnetic beads (Miltenyi Biotec, Teterow, Germany) for 20 min. Cells were then separated into CD45+ and CD45− cells using magnetic columns (Miltenyi Biotec) and plated onto fibronectin-coated chamber slides (BD Biosciences) in EGM-2 medium supplemented with 50 ng/ml vascular endothelial cell growth factor (VEGF, Lonza, Walkersville, MD, USA). After 14 days of induction, cells were incubated with DiI-ac-LDL (Invitrogen) as previously described, or fixed and stained with anti-von Willebrand factor (vWF) (Santa Cruz Biotechnology) or anti-endothelial nitric oxide synthase (eNOs) (BD Biosciences) antibodies and visualized with an Alexa Fluor 488-conjugated secondary antibody. Three representative fields were recorded and the positive cells were calculated by Image-Pro Plus software (Media Cybernetics, Rockville, MD, USA).
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