indole production was measured as described in [46 (link)]. To this end, 0.4 mL of cell-free culture supernatant was treated with the same volume of 20% trichloroacetic acid, mixed, incubated 10 min on ice and cleared by centrifugation. Then 0.4 mL of the treated supernatant was reacted with the same volume of Kovac’s reagent (Sigma-Aldrich). indole production was estimated using a standard curve of commercial indole (Sigma-Aldrich) and reported as μM indole normalized by the culture OD600. Hemolysin activity on sheep red blood cells was conducted as described in [47 ] except that a unit of enzyme activity was defined as the amount of enzyme causing an increase in 0.01 optical density units at 570 nm per h at 37°C. Biofilm formation was measured by the crystal violet staining method and expressed as optical density at 570 nM [33 (link), 34 (link)].
Indole
Manufactured by Merck Group
Sourced in United States, Germany, Canada
Indole is a heterocyclic organic compound used in various laboratory applications. It serves as a core chemical structure in a range of compounds and is utilized in analytical and research procedures.
Automatically generated - may contain errors
Lab products found in correlation
Phenol, by Merck Group (6 mentions)
Linalool, by Merck Group (6 mentions)
Hexanal, by Merck Group (5 mentions)
P cresol, by Merck Group (5 mentions)
Benzaldehyde, by Merck Group (5 mentions)
Methanol, by Merck Group (5 mentions)
Tryptophan, by Merck Group (4 mentions)
Phenylethyl alcohol, by Merck Group (4 mentions)
Heptanal, by Merck Group (4 mentions)
Decanal, by Merck Group (4 mentions)
6 methyl 5 hepten 2 one, by Merck Group (4 mentions)
Benzyl alcohol, by Merck Group (4 mentions)
Camphene, by Merck Group (4 mentions)
Octanal, by Merck Group (4 mentions)
Nonanal, by Merck Group (4 mentions)
3 methylindole, by Merck Group (4 mentions)
Acetophenone, by Merck Group (4 mentions)
Ethanol, by Merck Group (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Benzeneacetaldehyde, by Merck Group (3 mentions)
92 protocols using indole
1
Measuring Indole, Hemolysin, and Biofilm
2
Oral Gavage Probiotic Modulation
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All mice were treated by oral gavage daily thorough out the course of the experiments. Mice received indole by gavage (20 mg/kg/dose or ~200 μL of 17 mM indole; Sigma Aldrich, St. Louis, MO) dissolved in sterile water warmed to 55 °C. All probiotics (Bifidobacterium bifidum, Bifidobacterium lactis, Lactobacillus plantarum, Lactobacillus reuteri, and Lactobacillus rhamnosus) were obtained from DuPont™ Danisco® FloraFIT® probiotics as lyophilized powder at a known CFU/gram concentration. Each probiotic was weight and aliquoted into 1 × 108 CFU aliquots. Probiotics or heat-killed probiotics were resuspended in PBS and mice were gavaged with 5 × 108 CFU/dose. Probiotic cocktails were heat-killed by incubating at 100 °C for 15 min, bacterial viability was then confirmed by plating. No viable organisms were recovered from the heat-killed probiotics. Mice received CH-223191 by gavage (10 mg/kg/dose or ~200 µL of 3 mM CH-223191; Sigma Aldrich) dissolved in sterile corn oil. Alcohol-fed and pair-fed mice were gavage daily with vehicle (PBS). A separated cohort of control mice was given corn-oil or water daily to ensure no effects were seen with the different vehicles. No changes in host defense or bacterial burden were seen in control mice given corn-oil or water.
3
Epithelial Cell Culture Protocol
4
Catalase and Indole Derivatives Assay
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Bovine liver catalase, indigo, indole, and indole derivatives were purchased from Sigma-Aldrich. All other chemicals were analytical grade and obtained from Sigma-Aldrich or Merck. NADPH and NADP + were purchased from Oriental Yeast Co. LTD.
5
Indole's Influence on Hydrogen Production
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To evaluate the effect of indole on hydrogen production, pre-inocula were grown aerobically overnight in LB medium at 37°C, shaken at 150 rpm. Anaerobic experiments were performed in 120 mL serological bottles containing 110 mL of HP medium. Bottles were closed with 20 mm butyl septum and an aluminum crimp seal (Supelco) using an E-Z crimper (Wheaton). The cultures were started at pH 7.2 with an initial OD 600 of 0.3 and incubated at 37°C in a water bath shaking at 175 rpm.
Concentrations of indole (Sigma) between 0.5 to 8 mM were assessed, for this the required volume of a 2 M indole stock was added to the serological bottles using a sterile syringe. Control cultures were grown in HP medium plus 400 µL of ethanol (ethanol was used as the solvent for the stock indole solution). Samples of 1 mL taken at various times during fermentation were centrifuged at 13,000 rpm and stored at -20°C until analyzed as described below.
Concentrations of indole (Sigma) between 0.5 to 8 mM were assessed, for this the required volume of a 2 M indole stock was added to the serological bottles using a sterile syringe. Control cultures were grown in HP medium plus 400 µL of ethanol (ethanol was used as the solvent for the stock indole solution). Samples of 1 mL taken at various times during fermentation were centrifuged at 13,000 rpm and stored at -20°C until analyzed as described below.
6
Synthesis of Amino Acid Derivatives
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l -Tyrosine methyl ester hydrochloride (TyrMe), l -tryptophan methyl ester hydrochloride (TrpMe), acetonitrile, 2-propanol (all spectroscopic grade), phenol, 4-cyanophenol, 4-methoxyphenol, indole, methylviologen and deuterated water were purchased from Sigma Aldrich or Tedia. Water was Milli-Q grade. N-Acetyl l -tryptophan methyl ester (NATrpME)66 (link) and N-acetyl l -tyrosine methyl ester (NATyrME)67 (link) were prepared by literature procedures. The naphthoquinone derivatives were prepared as described in the literature.60,61,68 (link)
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7
Biochemical Assays for Tryptophan, Indole, and Albumin
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Assays for Tryptophan, indole (Sigma Aldrich, St. Louis, MO) and albumin (Sigma Aldrich, St. Louis, MO) levels were performed using the commercially available kits according to manufacturer's protocol.
8
Characterization of Aroma Compounds
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Standard 2‐hexanone, hexanal, 1H‐1‐ethyl‐pyrrole, 2‐hexenal, cis‐3‐hexen‐1‐ol, heptanal, 6‐methyl‐5‐hepten‐2‐one, 2‐pentyl‐furan, 2‐ethyl‐1‐hexanol, benzeneacetaldehyde, cis‐linalool oxide, trans‐linalool oxide, linalool, 3‐octen‐2‐ol, phenylethyl alcohol, menthol, α‐terpineol, safranal, decanal, camphene, geraniol and indole were purchased from Sigma Co. Ltd. Benzyl alcohol, trans‐β‐damascenone, trans‐α‐ionone, cis‐geranylacetone, trans‐β‐ionone, 2,4‐ditert‐butylphenol, cedrol, and caryophyllene oxide were purchased from Alfa Aesar Co. Ltd. Standard chemical series of C8–C20 alkanes that were used to determine the liner retention index (RI) and the internal standard cyclohexanone were obtained from Sigma Co. Ltd. Cis‐pyranoid‐linalool oxide and trans‐pyranoid‐linalool oxide were purchased from Sinopharm Chemical Reagent Co. Ltd.
9
Gut Microbiome Modulation Protocol
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The main regents used in this study, Trp, indole, IPA, IAld, Ficz, CH223191, metronidazole, ampicillin, vancomycin, and neomycin sulfate, were purchased from Sigma-Aldrich (St. Louis, MO, USA). The specific primary antibodies of AhR and Cyp1a1 were bought from Affinity Biosciences (OH, USA). Phosphorylation (p-) of p65 and IκB, p65, IκB, and β-actin were obtained from Cell Signaling Technology (CST; Boston, USA). The occludin and claudin-3 were bought from Bioss (Beijing, China). MPO activity assay kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TNF-α and IL-1β enzyme linked immunosorbent assay (ELISA) kits were bought from Biolegend, Inc. (San Diego, CA, USA). indole and IAld ELISA kits were bought from Hnybio (Shanghai, China), and the IPA ELISA kit was obtained from Shyqbio (Shanghai, China). L. reuteri CNCM I-5022 was obtained from the Collection Nationale de Cultures de Microorganisms (CNCM) of the Institut Pasteur. E.coli CVCC1418 was obtained from the China Veterinary Culture Collection Center (CVCC).
10
Analytical Standards for Aroma Profiling
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The standards 2-hexanone, benzeneacetaldehyde, linalool, linalool oxides, phenylethyl alcohol, 4-oxoisophorone, menthol, α-terpineol, safranal, geraniol, indole, and 2-ethyl-3-methylpyrazine were purchased from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA). The standards, 1-ethylpyrrole, benzyl alcohol, 1,2-dimethoxybenzene, 3,4-dimethoxytoluene, 1,2,3-trimethoxybenzene, 1,2,4-trimethoxybenzene, trans-β-ionone, 2,4-ditert- butylphenol, and dihydroactinidiolide were obtained from Alfa Aesar Co., Ltd. (Heysham, Lancashire, UK). A standard series of C8–C20 alkanes were used for retention index (RI) determination, and the internal standard cyclohexanone were purchased from Sigma-Aldrich Co., Ltd. (St. Louis, Mo, USA). Other reagents were all of analytical grade and obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
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