FBG, FINS, TG, TC, and FFA were measured. TG, TC and FFA were determined using an automatic analyzer (Roche, Basel, Switzerland). Serum insulin levels were measured by enzyme-linked immunosorbent assay (Jiancheng, Nanjing, China). The aorta was separated and fixed by formalin, and HE staining was performed to observe the histomorphology. The expression and distribution of NOX4 and VCAM-1 in aorta were detected by immunohistochemical staining. Cell apoptosis were detected by TUNEL assays.
Enzyme linked immunosorbent assay
Manufactured by Nanjing Jiancheng
Sourced in China
The enzyme-linked immunosorbent assay (ELISA) is a laboratory technique used to detect and measure specific proteins, peptides, antibodies, or hormones in a sample. It is a widely used analytical tool in various fields, including immunology, biochemistry, and molecular biology. The core function of ELISA is to quantify the presence and concentration of target analytes in a sample by using antibodies or antigens that are linked to an enzyme, which catalyzes a color-producing reaction. This process allows for the detection and measurement of the target analyte with high sensitivity and specificity.
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Lab products found in correlation
8 protocols using enzyme linked immunosorbent assay
1
Aortic Histomorphology and Oxidative Stress
2
Serum Biochemical Analysis in Rats
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Blood samples were taken from the orbital sinus of animals in each group (7, 14, 21, and 28 days) under light ether anesthesia. Blood was collected and kept for 1 h at room temperature for clotting. Serum was separated by centrifuging the blood sample at 3000 rpm for 20 min. The biochemical parameters, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), ammonia, bilirubin, and total protein content in the serum of each rat, were evaluated using enzyme-linked immunosorbent assay according to the kit protocols (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
3
Fasting Biochemical Markers Measurement
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After the rats were fasted overnight, blood samples were obtained from the jugular vein. Total cholesterol (TC), triglyceride (TG), FBG, and free fatty acid (FFA) levels were measured using the corresponding detection kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Fasting insulin (FINS) was measured using enzyme-linked immunosorbent assay (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The insulin sensitivity index (ISI) was calculated using the following formula: ISI = ln [([FBG] × [FINS])−1].
4
Zebrafish Immune Protein Expression
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Zebrafish embryos (n = 150) were homogenized in precooled dilution buffer then centrifuged at 2500 rpm for 10 min at 4 °C; the supernatant was taken for measurements. The expression of immune-related proteins (interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), nuclear factor-κB (NF-κB), Toll-like receptor 4 (TLR4)) in zebrafish embryos was detected using the enzyme-linked immunosorbent assay (Nanjing Jiancheng Institute of Biological Engineering, Nanjing, China) in strict accordance with the manufacturer’s instructions. All protein indexes were measured in triplicate using a microplate reader.
5
Oxidative Stress Markers in Follicular Fluid
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The GCs were isolated from the aspirated FF in all patients using gradient centrifugation. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the FF were measured using thiobarbituric acid and the chemiluminescence technique, respectively. In the FF, 8-OHdG was quantitatively detected by enzyme-linked immunosorbent assay (Jiancheng, Nanjing, China). The evaluations were repeated three times.
6
Mucosal Antioxidant and Immune Markers
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Approximately 0.3-g mucosal samples were minced and placed in ice-cold 154 mmol/L sterile sodium chloride solution (1:9, wt/vol) containing protease inhibitors and then homogenized with an Ultra-Turrax homogenizer (Tekmar, Cincinnati, OH, USA) at a high speed for 30 s. Samples were spun at 4450 × g for 15 min at 4°C. The supernatant was aliquoted and stored at −80°C for analysis.
The activity of total superoxide dismutase (T-SOD) and the concentrations of total protein, malondialdehyde (MDA), and reduced glutathione (GSH) were quantified using colorimetry kits from the Nanjing Jiancheng Institute of Bioengineering (Nanjing, Jiangsu, China), as per the manufacturer's instructions. The contents of secretory immunoglobulin A (SIgA), immunoglobulin G (IgG), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the mucosal samples were measured by enzyme-linked immunosorbent assay (Nanjing Jiancheng Bioengineering Institute). All results were normalized against total protein concentration in each sample for inter-sample comparison.
The activity of total superoxide dismutase (T-SOD) and the concentrations of total protein, malondialdehyde (MDA), and reduced glutathione (GSH) were quantified using colorimetry kits from the Nanjing Jiancheng Institute of Bioengineering (Nanjing, Jiangsu, China), as per the manufacturer's instructions. The contents of secretory immunoglobulin A (SIgA), immunoglobulin G (IgG), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the mucosal samples were measured by enzyme-linked immunosorbent assay (Nanjing Jiancheng Bioengineering Institute). All results were normalized against total protein concentration in each sample for inter-sample comparison.
7
Fasted Rat Serum Lipid Analysis
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After rats fasted overnight, serum triglycerides (TG) and total cholesterol (TC) were determined by use of an automatic analyzer (Roche, Basel, Switzerland). Serum level of insulin was measured by enzyme-linked immunosorbent assay (Jiancheng, Nanjing, China).
8
Serum Cytokine Quantification
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The blood samples were taken from the heart after cervical dislocation. The serum was cleared from cellular components of the blood by centrifugation at 12,000 rpm for 10 min at 4 °C and stored at − 80 °C until use. IL-6 and TNF-α levels were measured using an enzyme-linked immunosorbent assay following the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). A blank hole without sample only adding chromogenic agent and terminated liquid were used for zero. Optical density (OD) at 450 nm was read using a Microplate Reader (Corona Electric, Hitachinaka, Japan).
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