Plasma alcohol concentrations were measured using an Ethanol Assay Kit (Sigma-Aldrich, St. Louis, MO, USA). Plasma enzyme activities of AST and ALT were colorimetrically determined by the corresponding commercial assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), according to the manufacturer’s instructions. Plasma levels of free fatty acid, glucose, and insulin were measured by a Free Fatty Acid Quantification Colorimetric Kit (BioVision, Milpitas, CA, USA), a glucose assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and a Mouse Insulin ELISA kit (Westang Biological Technology Co., Ltd, Shanghai, China), respectively, according to the manufacturers’ instructions.
Ethanol assay kit
Manufactured by Merck Group
Sourced in United States
The Ethanol Assay Kit is a lab equipment product designed to measure the concentration of ethanol in various samples. It provides a quantitative analysis of ethanol levels through a colorimetric reaction. The kit includes the necessary reagents and protocols to perform the assay.
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Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Free fatty acid quantification colorimetric kit, by Abcam (1 mentions)
Glucose assay kit, by Nanjing Jiancheng (1 mentions)
Vacutainer, by BD (1 mentions)
Alt activity assay kit, by Merck Group (1 mentions)
Rat mouse insulin elisa kit, by Merck Group (1 mentions)
Infinite m1000 pro multimode plate reader, by Tecan (1 mentions)
Male sprague dawley rats, by Orient Bio (1 mentions)
Reflotron, by Roche (1 mentions)
Reflotron plus, by Roche (1 mentions)
Edta collection tubes, by Greiner (1 mentions)
Genios pro, by Tecan (1 mentions)
Dc protein assay, by Bio-Rad (1 mentions)
Serum separator tube, by BD (1 mentions)
Lobind tubes, by Sarstedt (1 mentions)
17 protocols using ethanol assay kit
1
Plasma Biomarker Profiling Protocol
2
In vitro Ethanol Exposure Methodology
3
Ethanol Concentration Colorimetric Assay
4
Alcohol Blood Sampling and Analysis
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One milliliter of maternal blood was collected in heparin BD Vacutainer® by cardiac puncture 45 min after the administration of alcohol (either at Med or Bin doses) or maltodextrin. Samples were maintained for five minutes at room temperature and centrifuged at 1750× g for 20 min at 4 °C. Blood alcohol concentrations (BAC) were determined by measuring absorbance at 570 nm in serum samples using the Ethanol Assay Kit (MAK076, Sigma-Aldrich, Sant Louis, MO, USA) following the indications of the manufacturer.
5
Serum Biomarkers in Ethanol Study
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The baseline blood samples were taken 2 weeks prior to starting the control or ethanol diet. Serum was collected from the terminal blood of the mice at sacrifice. For the analysis of serum acetate at the 2-week time point of the ethanol diet, 20 μl of hind-limb venous blood was collected into a capillary. The isolated serum together with the baseline and sacrifice serum samples was analyzed with the acetate colorimetric assay kit (MAK086, Sigma-Aldrich). The ALT activity assay kit (MAK052, Sigma-Aldrich) was used to determine serum ALT levels. Serum total cholesterol, HDL cholesterol and triglyceride levels were determined by an enzymatic method (Roche Diagnostics), and LDL + VLDL cholesterol values were calculated using the Friedewald equation [16 (link)]. Serum insulin levels were determined with the Rat/Mouse Insulin ELISA kit (EZRMI-13 K, Millipore). Blood glucose concentrations were measured with a glucometer and the homeostatic model assessment-insulin resistance (HOMA-IR) index was calculated from the blood glucose and serum insulin values. Blood ethanol levels were determined with the Ethanol Assay Kit (Sigma Aldrich, MAK076-1KT) by a coupled enzyme reaction that results in a colorimetric product. The absorbances of the colorimetric products were determined with the Infinite M1000 Pro Multimode Plate Reader (Tecan).
6
Ethanol Assay of Blood Samples
7
Ethanol-Induced Liver Damage Assessment
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For in vivo assessment, the WiKim0168 and WiKim0172 strains were prepared by recovering the culture from a mixed medium, freeze-drying it in combination with a cryoprotectant, and storing it at 4°C as a freeze-dried powder until use. The care and study of the laboratory mice followed the protocols of the Institute Animal Care and Use Committee for the World Institute of Kimchi (WIKIM IACUC 202203). Male Sprague–Dawley rats (six weeks old; OrientBio, Republic of Korea) were fasted for 8 h, whereafter they were orally administered 25%ethanol and the LAB strains (1 × 1010 CFU). Blood samples were collected from rat tails at 30, 60, 180, and 300 min post-administration, centrifuged to separate the serum, and the ethanol concentration was determined using an Ethanol Assay Kit (MAK076; Sigma-Aldrich, USA). To assess liver damage, liver function was analyzed by monitoring the indicators of serum lipid metabolism using DRI-CHEM NX500 (Fujifilm Global, Japan) analyses for aspartate aminotransferase (AST) activity, total cholesterol, and triglycerides.
8
Evaluating Lb NOX Effects on Ethanol Production
9
Ethanol Absorption Kinetics in Mice
10
Ethanol Production in Bacillus anthracis
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B. anthracis WT cells were grown at 30 °C in LB medium overnight and subcultured at a 1:100 ratio into fresh LB medium amended with 0 or 50 µM ‘205. After 6 h of growth at 37 °C, cells were harvested by centrifugation and the spent media were filtered through a 0.22-µm membrane filter. The ethanol content was quantified using an ethanol assay kit (cat# MAK076, Sigma-Aldrich, MO) according to the manufacturer’s instructions. All samples and standards were run in duplicate and fluorescence intensity (λex = 535 nm and λem = 587 nm) was monitored for 30 min. Experiments were carried out three times with three biological replicates each time.
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