The feces of mice in the N + N and Abs+N groups were vortex mixed, respectively, with cysteine (Solarbio, China) and Na2S (Solarbio, China) in corresponding centrifuge tubes (500 mg feces, 2.5 mg cysteine, and 1 mg Na2S in 3 mL bacteria-free PBS). Then, the mixture for each group was centrifuged (500 r·min−1, 1 min) and the supernatant solution was collected. Mice in the FMT-N and FMT-Abs groups received intragastric administration of the supernatant solution from the N + N and Abs+N groups, respectively, (100 μL per mouse).
Cysteine
Manufactured by Solarbio
Sourced in China
Cysteine is a non-essential amino acid that plays a crucial role in the structure and function of proteins. It is a sulfur-containing amino acid that can form disulfide bridges, which contribute to the stability and tertiary structure of proteins. Cysteine is commonly used in various biochemical and biological applications, including protein purification, enzymatic assays, and cell culture media formulations.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Cellobiose, by Maclin Biochemical Technology (1 mentions)
Maltose, by Solarbio (1 mentions)
Glutathione (gsh), by Beyotime (1 mentions)
Lysozyme, by Solarbio (1 mentions)
Bull serum albumin bsa, by Solarbio (1 mentions)
Milli q plus system, by Merck Group (1 mentions)
Trypsin, by Solarbio (1 mentions)
Pepsin, by Solarbio (1 mentions)
Pepsin, by Yuanye Bio-Technology (1 mentions)
Trypsin, by Yuanye Bio-Technology (1 mentions)
L rhamnose, by Yuanye Bio-Technology (1 mentions)
D glucose, by Yuanye Bio-Technology (1 mentions)
L arabinose, by Yuanye Bio-Technology (1 mentions)
D mannose, by Yuanye Bio-Technology (1 mentions)
α amylase, by Yuanye Bio-Technology (1 mentions)
Pancreatin, by Merck Group (1 mentions)
Tween 80, by Solarbio (1 mentions)
Pig bile salt, by Solarbio (1 mentions)
Feso4, by Merck Group (1 mentions)
8 protocols using cysteine
1
Fecal Microbiome Transplant Mice
2
Anaerobic Gut Bacteria Cultivation Protocol
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Roseburia intestinalis L1-82 (DSMZ14610) was purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany). Bacteroides fragilis (ATCC 25285) and Bacteroides vulgatus were obtained from Ningbo Mingzhou Biotechnology Co., Ltd. (Zhejiang, China). The above bacteria were cultured under anaerobic conditions at 37°C. An anaerobic culture environment was created using Atmosphere Generation Systems (AnaeroJar™ ASSEMBLY and AnaeroGen™, OXOID, Thermo Fisher Scientific). The culture medium (Miquel et al., 2015 (link)) was brain heart infusion (BHI) containing 0.5% yeast extract (OXOID), 1 mg/ml cellobiose (Macklin, China), 1 mg/ml maltose (Solarbio, China), 0.5 mg/ml cellobiose (Macklin, China), and 0.5 mg/ml cysteine (Solarbio, China). We established a standard curve to convert the absorbance values at 600 nm (OD600) to CFU values (Supplementary Figure S1 ). Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Salmonella typhimurium CMCC 50115, Bacillus subtilis DSM 1088, and Bacillus cereus ATCC 11778 were used as controls. These strains were cultured in nutrient broth at 37°C in an aerobic environment.
3
Cysteine and GSH Oxidation Assay
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Totally, 20 μl 10 mM Cysteine (Solarbio, Beijing, China) in PBS or 20 μl 10 mM GSH (Beyotime Biotechnology, Shanghai, China) in PBS was incubated with 60 μl of 0, 0.05, 0.25, 0.5, or 1 mM NaIO3 in PBS for 10 min at 37 °C. The oxidation of the thiol group in Cysteine or GSH was monitored with a Cysteine assay kit as indicated above.
4
Biochemical Assay Protocol Development
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Sulfamide and m-phenylenediamine (m-PD) were obtained from J&K Scientific Ltd. (Beijing, China). Sodium pyrophosphate and amino acids were obtained from Sangon Biotechnology Co., Ltd. (Shanghai, China). Alkaline phosphatase (ALP, EC 3.1.3.1, 2 kU) was purchased from Sigma (USA). Glucose oxidase (GOX), thrombin, trypsin, pepsin, lysozyme, cysteine and bull serum albumin (BSA) were purchased from Solarbio (Beijing, China). Other reagents were purchased from Tianjing Chemical Reagents Company (Tianjing, China). All chemicals were used as received without further purification. HEPES buffer (10 mM, pH 7.4) was used throughout the experiments. Ultrapure water was obtained from a Milli-Q Plus system (Millipore, Bedford, MA, USA) and used in all experiments.
5
Extraction and Analysis of C. axillaris Fruit
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C. axillaris fruit was purchased from Ganzhou, Jiangxi Province, China. α-amylase, pepsin, trypsin and monosaccharide standards (L-fucose, L-Fuc; L-rhamnose, L-Rha; L-arabinose, L-Ara; D-glucosamine, D-GlcN; D-glactose, D-Gal; D-glucose, D-Glc; L-xylose, L-Xyl; D-mannose, D-Man; L-fructose, L-Fru; D-galacturonic acid, D-GalA; D-guluronic acid, D-GulA) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd (Shanghai, China). Pancreatin was purchased from Sigma Chemical Co. (St. Louis, MO, USA). Inulin, resazurin and fatty acid standard (acetic acid, propionic acid, i-butyric acid, n-butyric acid, i-valeric acid, n-valeric acid, n-hexanoic acid) were bought from Shanghai Aladdin Biochemical Technology Co., Ltd (Shanghai, China). Cysteine, heme chloride, pig bile salt and Tween-80 were from Beijing Solarbio Science & Technology Co.,Ltd. (Beijing, China). Dextran standards (Mw 1152, 5000, 11600, 23800, 48600, 80900, 148000, 273000 Da) were bought from Sigma Chemical Co. Ltd. (St. Louis, MO, USA).
6
HSCCC Separation and Antioxidant Assay of B. simaoensis
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The solvents used for the HSCCC separation and antioxidant assay were analytical grade, and were purchased from Tianjing Chemical Factory (Tianjing, China). Acetontrile was HPLC grade and purchased from Siyou Special Reagent Factory (Tianjin, China). The PBS, liver microsomes, and cysteine were bought from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). The FeSO4 and thiobarbituric acid were obtained from Sigma (St. Louis, MO, USA). The whole plants of B. simaoensis were collected in Yunan, China, and identified by Dr. Jia Li (College of Pharmacy, Shandong University of Traditional Chinese Medicine).
7
Protein Delivery and Imaging in Cells
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BioPORTER® QuikEase™ Protein Delivery Kit, pluronic F127, progesterone, IP3R1 antibody (rabbit), and fluorescein isothiocyanate-labeled peanut (Pisum sativum) agglutinin (FITC-PSA) were purchased from Sigma-Aldrich Chemical Co. (St Louis, MO, USA). Earle's balanced salt solution (EBSS), phosphate-buffered saline (PBS), and penicillin–streptomycin were obtained from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Sodium pyruvate and triethanolamine were purchased from Amresco, Inc. (Solon, OH, USA). Hoechst 33258 and polyvinylpyrrolidone-40 (PVP-40) were obtained from Shanghai Maokang Biotechnology Co., Ltd. (Shanghai, China). Bovine serum albumin (BSA) and glutaraldehyde were purchased from Sangon Biotech Co. (Shanghai, China). Cysteine and chlortetracycline (CTC) were obtained from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). Rabbit IgG was purchased from Abcam Plc. (Cambridge, MA, USA). Fluo-3AM was obtained from Beyotime Biotechnology (Shanghai, China). The calcium ionophore A23187 was obtained from 4A Biotech Co., Ltd. IP3 was purchased from MedChem Express LLC (Monmouth Junction, NJ, USA). The FITC labeling kit was obtained from Elabscience Biotechnology Co. (Wuhan, China). All other chemicals were reagent grade.
8
Synthesis and Characterization of Organic and Inorganic Compounds
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ATP, p-toluidine, 5-ASA, 3-aminosalicylic acid (3-ASA), 4-aminosalicylic acid (4-ASA) were obtained in Shanghai Perfemiker Material Tech Co., Ltd (Shanghai, China, http://www.canspecsci.com ). Fe(NO3)3, CuCl2, HgCl2, NiCl2, CdCl2, CrCl3, ZnCl2, CaCl2, Pb(NO3)2, AgNO3, Co(NO3)2·6H2O, Mn(Ac)2, MgCl2, BaCl2 and Bi(NO3)3·5H2O were purchased from Fluka (Buchs, Switzerland, http://www.fluka.org/fluka.php ). p-Nitrochlorobenzene, o-nitrobenzaldehyde, p-nitrotoluene, salicylic acid, benzidine, p-chlorophenol, acetanilide, o-aminophenol, alanine, leucine, phenylalanine, glutamic acid, asparagine, arginine, tyrosine, histidine, serine, glycine, valine, tryptophan, lysine, glutathione, cysteine, melamine and ethanol were purchased in Beijing Solarbio Reagent Co., Ltd (Beijing, China, http://www.solarbio.com ). Unless otherwise specified, all chemicals from commercial sources are analytically pure and can be used directly without further purification. All solutions were prepared and diluted with ultrapure water from the MillI-Q water purification system.
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