Penises were fixed with 4% formaldehyde supplemented with 0.002% picric acid for 4 hours at room temperature and were then dehydrated in 30% sucrose PBS overnight at 4 °C. Fixed tissues were embedded in optimal cutting temperature (OCT) compound (Leica) and cut into 10-μm sections. Sections were blocked with 0.1% Triton X-100/0.1% Tween 20/3% nonimmune goat serum in PBS for 1 hour at room temperature Sections were incubated with primary antibodies (anti-MYPT1 (1:200, Cat. No. 22117-1-AP, Proteintech); anti-smooth muscle α-actin (1:200, Cat. No. ab7817, Abcam); anti-eNOS (1:200, Cat. BS3571, BioRad); antidesmin (1:200, Cat. No. 21404-1-AP, Proteintech); anti-SMHHC (1:200, Cat. No. 16520-1-AP, Proteintech), anti-CD31 (1:200, Cat.No.550274, BD Biosciences) overnight at 4 °C and then with fluorescent secondary antibodies (Invitrogen) for 2 hours at room temperature. Fluorescence staining was examined under a confocal microscope (Olympus).
Anti enos
Manufactured by Bio-Rad
Sourced in United States
Anti-eNOS is a laboratory reagent designed to detect the endothelial nitric oxide synthase (eNOS) protein. eNOS is an enzyme responsible for the production of nitric oxide, which plays a crucial role in various physiological processes. Anti-eNOS is used in research applications to identify and quantify the presence of eNOS in biological samples.
Automatically generated - may contain errors
2 protocols using anti enos
1
Immunohistochemical analysis of penile tissue
2
Placental Protein Expression Analysis
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Placenta samples (100 mg) were frozen with liquid nitrogen, homogenized in 1 ml RIPA buffer (high) (cat. no. R0010, Beijing Solarbio Science Technology Co., Ltd., Beijing, China) and then centrifuged at 12,000 × g for 10 min at 4°C, and the protein concentration was determined using NanoDrop-1000 spectrophotometer (Gene Co., Ltd., Beijing, China). Protein extracts were re-suspended in a 5X sample buffer and boiled in water for 15 min at 100°C. Total protein (100 µg) was subjected to 10% SDS-PAGE and transferred to nitrocellulose membranes (cat. no. 1620150; Bio-Rad Laboratories, Inc., Hercules, CA, USA), followed by incubation with the primary antibodies anti-eNOS (cat. no. sc-654; 1:1,000 dilution) and anti-GAPDH (cat. no. sc-25778; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight. Subsequently, membranes were incubated with the IRDye 800CW goat anti-rabbit immunoglobulin G (H+L) (Rockland Immunochemicals Inc., Pottstown, PA, USA) for 1 h at room temperature. Blots were visualized by enhanced chemiluminescence (Beijing Applygen Technologies, Beijing, China), and bands were analyzed by using ImageJ software (vision 1.37; NIH, Bethesda, MD, USA). The protein contents were normalized to that of GADPH.
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