Animals were anesthetized using 3% pentobarbital sodium 30 mg/kg injected via the marginal ear vein. The cortex and hippocampus of the rabbits were separated after the brains were extracted. A 10% w/v rabbit cortex and hippocampus homogenate were made in pre-chilled physiological saline using an IKA ULTRA-TURRAX homogenizer (IKA®-werke Gmbh & Co., KG, German), the obtained homogenate was centrifuged at 4500 rpm, 4 °C for 10 min. The supernatant was extract and the Aβ1–42 level was assayed immediately using homologous specific ELISA kits (Nanjing Jiancheng Bioengineering Institute, China), according to the manufacturer’s instructions. Protein concentration was determined by Coomassie brilliant blue method (Nanjing Jiancheng Bioengineering Institute, China).
Coomassie brilliant blue method
Manufactured by Nanjing Jiancheng
Sourced in China
Coomassie brilliant blue method is a colorimetric assay used to quantify the total protein concentration in a sample. It relies on the binding of the Coomassie brilliant blue dye to proteins, resulting in a color change that can be measured spectrophotometrically.
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Lab products found in correlation
3 protocols using coomassie brilliant blue method
1
Alzheimer's Biomarker Quantification
2
Lung Oxidative Stress Biomarkers
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Lung homogenate (10%) was prepared with an ultrasonic cell grinder. MDA (malondialdehyde) and NO (nitric oxide) levels in the lung homogenate were detected with commercially available kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, Jiangsu, China) according to the manufacturer’s instructions. Protein contents were determined with the Coomassie brilliant blue method (Nanjing Jiancheng Bioengineering Institute). The TNF-α level was assessed with radioimmunoassay (RIA) by the Beijing Yinghua Biotechnical Institute, Beijing, China. Peroxynitrite (ONOO-) content in the lung tissue was analyzed with ELISA by the Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China). At least three independent measurements were performed for each index.
3
Biochemical Assays in Clam Tissues
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Prior to the biochemical measurements, six C. uminea of each treatment group were rinsed with Milli-Q® water, and the organs (gills and digestive glands) were dissected using clean forceps and a scalpel. The entire dissection was carried out on an ice plate. Afterwards, the soft tissues were homogenized using ultra-sonication in pH 7.4, 0.1 mol/L PBS buffer solution (137mM NaCl, 2.7mM KCl, 10mM Na 2 HPO 4 , 1.8mM KH 2 PO 4 ) under an ice water bath. Afterwards, the samples were centrifuged (4°C) at 8000 × g for 15 minutes, and the supernatant was saved for further analysis.
The biochemical indices employed (content of MDA, the activity of antioxidant enzymes (SOD, CAT, GSH-Px, GR, and GST), and the content of GSH) were measured following the instructions of the respective assay kits (Nanjing Jiancheng Bioengineering Institute, China). The results from biochemical analyses were normalized to sample total protein. A protein assay kit (Coomassie brilliant blue method, Nanjing Jiancheng Bioengineering Institute, China) was used for the analysis of protein content in all of the samples.
The biochemical indices employed (content of MDA, the activity of antioxidant enzymes (SOD, CAT, GSH-Px, GR, and GST), and the content of GSH) were measured following the instructions of the respective assay kits (Nanjing Jiancheng Bioengineering Institute, China). The results from biochemical analyses were normalized to sample total protein. A protein assay kit (Coomassie brilliant blue method, Nanjing Jiancheng Bioengineering Institute, China) was used for the analysis of protein content in all of the samples.
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