The body weight was measured weekly. Samples were collected every 24 h from rats housed in metabolic cages for 0, 2, 5, 7, 9 and 10 weeks with access to drinking water only. The urinary protein and albumin excretion were measured using the Coomassie Blue method (Jiancheng, Nanjing) or an ELISA kit according to the manufacturer’s instructions. The urinary N-acetyl-β-D-glycosaminidase (NAG) levels was measured using an ELISA kit (Jiancheng, Nanjing). Blood samples were collected on weeks 0, 2, 5, 7, 9 and 10 (at death) from the inner canthus or heart after sacrifice in the 10th week to assess the changes in biochemical parameters (Hitachi, Tokyo, Japan).
Coomassie blue method
Manufactured by Nanjing Jiancheng
Sourced in China
The Coomassie blue method is a laboratory technique used for the quantitative determination of protein concentrations. It involves the binding of a dye, Coomassie Brilliant Blue, to proteins, which results in a color change that can be measured using a spectrophotometer. The intensity of the color is proportional to the amount of protein present in the sample.
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Lab products found in correlation
3 protocols using coomassie blue method
1
Urinary Biomarkers in Rat Metabolic Cages
2
Quantifying Mucosal Immunity and Cytokines in Porcine Samples
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Accordance with the manufacturer’s instructions, porcine-specific ELISA kits were used to quantify circulating immunoglobulin (IgA and IgG) (Bethyl Laboratories, Montgomery, TX). Immunoglobulin (sIgA and IgG) in the mucosa and cytokines (IL-8, IL-10, TNF-α and IFN-γ) in the serum and mucosa were measured using commercially available ELISA kits (Beijing 4A BiotechCo., Ltd., Beijing, China) in accordance with the manufacturer’s instructions and run in duplicates. Before the assays, the mucosa samples were vortexed in PBS (1:10, w/w) for 60 s and centrifuged at 5,000×g for at 4 °C, and the supernatant was obtained and used for the determination of cytokines and Ig levels. The concentrations of each cytokine and Ig in the intestinal mucosa were standardized to the protein in each sample. Total protein content in the mucosa was determined with Coomassie blue method following the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
3
Assessing Renal Oxidative Stress Markers
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Renal tissue malondialdehyde (MDA) concentration was measured by the thiobarbituric acid (TBA) method. Amounts of lipid peroxides (LPO) were measured as the production of MDA. Absorbance was measured at 532 nm using a spectrometer (assay kit; Nanjing Jiancheng Bioengineering Institute). Superoxide dismutase (SOD) activity in renal tissue was measured using a commercialized chemical assay kit (Nanjing Jiancheng Bioengineering Institute) by the xanthine oxidase method.
Absorbance was determined at 550 nm using a spectrometer. Glutathione peroxidase (GSH-Px) activity was determined by the colorimetric method using a GSH-Px kit (Nanjing Jiancheng Bioengineering Institute). Absorbance was determined at 412 nm using a spectrometer. All protein concentrations of renal tissue homogenate samples were determined with Coomassie blue method (assay kit; Nanjing Jiancheng Bioengineering Institute).
Absorbance was determined at 550 nm using a spectrometer. Glutathione peroxidase (GSH-Px) activity was determined by the colorimetric method using a GSH-Px kit (Nanjing Jiancheng Bioengineering Institute). Absorbance was determined at 412 nm using a spectrometer. All protein concentrations of renal tissue homogenate samples were determined with Coomassie blue method (assay kit; Nanjing Jiancheng Bioengineering Institute).
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