For HPLC analysis, esculin (EC, purity 98%) and esculetin (ECT, purity 98%) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Fraxin (FR, purity 98.9%) and formic acid (analytical reagent grade) were purchased from Merck KGaA (Darmstadt, Germany). For animal experiments, reserpine (purity 98%), esculin (purity 98%), esculetin (purity 98%), and fluoxetine (FXT, purity 98% in thin layer chromatography) were supplied by Sigma-Aldrich. Fraxin (purity 98%) was supplied by InterPharm Corporation (Koyang-si, Gyeonggi-do, South Korea). For anesthesia, tiletamine/zolazepam was supplied by Virbac (Zoletil 50; Cedex, France). For western blot analysis, actin antibody was supplied by Sigma- Aldrich. BDNF antibody was supplied by Abcam plc. (Cambridge, United Kingdom). CREB antibody and phosphorylated CREB (pCREB) antibody were supplied by Cell Signaling Technology (Danvers, MA, United STates). For immunofluorescence analysis, BDNF antibody was supplied by Abcam plc, pCREB antibody by Cell Signaling Technology, and NeuN antibody by Merck KGaA.
Esculin
Manufactured by Merck Group
Sourced in United States, Germany, Spain, Japan
Esculin is a laboratory reagent used as a substrate for the detection and identification of enzymes that can hydrolyze the glycosidic bond in this compound. It is commonly used in microbiological and biochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Esculetin, by Merck Group (6 mentions)
Coumarin, by Merck Group (4 mentions)
Scopoletin, by Merck Group (4 mentions)
Umbelliferone, by Merck Group (3 mentions)
Quercetin, by Merck Group (3 mentions)
Arbutin, by Merck Group (3 mentions)
Formic acid, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
4 methylumbelliferon, by Merck Group (2 mentions)
Formic acid, by Thermo Fisher Scientific (2 mentions)
Rosmarinic acid, by Merck Group (2 mentions)
Glycerol, by Merck Group (2 mentions)
Tween 20, by Merck Group (2 mentions)
P nitrophenyl β d glucopyranoside, by Merck Group (2 mentions)
Neun antibody, by Merck Group (1 mentions)
Fluoxetine, by Merck Group (1 mentions)
Pcreb antibody, by Cell Signaling Technology (1 mentions)
Reserpine, by Merck Group (1 mentions)
Bdnf antibody, by Abcam (1 mentions)
Creb antibody, by Cell Signaling Technology (1 mentions)
25 protocols using esculin
1
HPLC Analysis of Bioactive Compounds
2
Quantification of Cassava Phytochemicals
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Scopoletin and scopolin contents were measured as previously described [6 (link)]. One gram of cassava SRs was ground with 4 mL absolute ethanol, the extracts were centrifuged for 15 min at 10,000 rpm, and the supernatants were collected. The compounds in supernatants were identified by comparing their retention time with the standard samples as controls (scopolamine, scopolidine, esculin, and esculin; Sigma, Tokyo, Japan). Determination of the compositions and quantities of compounds in the supernatant was performed with the Agilent HPLC 1200 MS Q-TOF 6520 system. HPLC separation was performed according to the following conditions: a Zorbax Extend-C18 column (3.0 mm × 50 mm, 1.8 μm), with 98% H2O containing 20 mmol/L acetic amonium as solvent A and 2% ACN as solvent B, at a flow rate of 0.2 mL/min. An increasing gradient (v/v) of solvent B was used (time (min), A:B): 0, 98:2; 2, 95:5; 5, 90:10; 15, 85:15; 18, 45:55; and 20, 0:100. The mass spectrometric parameters were as follows: mass range 40~500 m/z (MS scan rate 1.4 spectra/s), positive scan, gas (N2) temperature 345 °C, gas flow 9 L/min, VCap 3400 V, Fragmentor 160 V, Skimmer 64 V, Octopole RF 750 V, and Ext Dyn Standard 2 GHz (1700).
3
Coumarin Compounds and Sorafenib Synergy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sorafenib (Nexavar, BAY 43-9006) (1 µM), osthole (150 µM), and hydroxycoumarins: 4-hydroxycoumarin (Sigma), umbelliferone, and esculin (Sigma) at the final concentrations of 200 µM were used in the experiments. The drugs were dissolved in DMSO (Sigma) to the final concentration not exceeding 0.01%. The doses were chosen based on previous studies [32 (link),33 (link)]. The cancer cells were treated with the coumarins or with Sorafenib separately or in combination for 24 h. As controls, T98G and MOGGCCM cells were incubated only with 0.01% of DMSO.
4
Quantifying Multispecies Biofilm Composition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plate count was established as the methodology to determine the bacterial growth of each of the strains in multispecies biofilms after incubation periods of 24, 48, and 72 h. To do so, the surfaces were washed twice with 3 mL of sterile distilled water to remove the unattached cells and then placed in a sterile flask containing 3.5 g of glass beads and 10 mL of peptone water. The samples were then vortexed for 90 s at 40 Hz to dislodge the attached cells from the surface for quantification [36 (link)].
The resulting suspension was decimally diluted in peptone water and transferred to a plate for its quantification. Since the biofilms consisted of two species, a culture medium was designed that enabled them to be differentiated. The media consisted of esculin, since L. gasicomitatum was observed to ferment the sugar, while P. fragi and L. reuteri did not. This enabled a medium composed of TSA, esculin (Sigma-Aldrich, Madrid, Spain), and iron citrate (Sigma-Aldrich, Madrid, Spain) to be developed, which turned the colonies of L. gasicomitatum black, making it easily distinguishable from the other two strains used. Differences were observed based on colony morphology. The plates were incubated at 30 °C for 48 h and then counted.
The resulting suspension was decimally diluted in peptone water and transferred to a plate for its quantification. Since the biofilms consisted of two species, a culture medium was designed that enabled them to be differentiated. The media consisted of esculin, since L. gasicomitatum was observed to ferment the sugar, while P. fragi and L. reuteri did not. This enabled a medium composed of TSA, esculin (Sigma-Aldrich, Madrid, Spain), and iron citrate (Sigma-Aldrich, Madrid, Spain) to be developed, which turned the colonies of L. gasicomitatum black, making it easily distinguishable from the other two strains used. Differences were observed based on colony morphology. The plates were incubated at 30 °C for 48 h and then counted.
5
Natural Compound Toxicity Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quercetin, escin, esculin and curcumin were purchased from Sigma-Aldrich. Toxin-containing media and control media were prepared by adding 100 µL of Quercetin, escin, esculin or curcumin dissolved in 100% EtOH or 100 µL of EtOH, respectively, to 5mL of molten yeast-sucrose media (5% w/v inactive yeast, 5% w/v sucrose, 1% w/v agar, 0.38% v/v propionic acid, 0.039% v/v orthophosphoric acid, 0.174% w/v Tegosept, 1.65% v/v EtOH) in each vial and mixing with a clean plastic rod. Media was stored at 4 °C for a maximum of three days before use.
6
Cinnamate and Coumarin Synthesis Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following acids were used: ferulic (Aldrich), coumaric (Sigma), cinnamic (Sigma), and caffeic (Fluka). coumarins used were: coumarin (Sigma), daphnetin (Sigma), esculetin (Sigma), esculin (Sigma), fraxetin (Extrasynthèse), fraxin (Extrasynthèse), isoscopoletin (Extrasynthèse), limetin (Herboreal), scoparon (Herboreal), 6-methoxycoumarin (Apin Chemicals), 7-methoxycoumarin (Herboreal), scopoletin (Herboreal), scopolin (Aktin Chemicals Inc.), umbelliferone (Extrasynthèse), skimmin (Aktin Chemicals Inc.), and 4-methylumbelliferon (Sigma). The CoA thiol esters of the cinnamates (cinnamoyl-CoA, p-coumaroyl-CoA, caffeoyl-CoA, and feruloyl-CoA) were enzymatically synthesized as described by Vialart et al. (2012) (link). P-coumarate and coenzyme A (CoA) were purchased from Sigma-Aldrich. Kanamycin, chloramphenicol, and isopropyl-β-d-thio-galactopyrannoside (IPTG) were purchased from Duchefa.
7
Quantification of Coumarin Standards
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The coumarins standards umbelliferone (purity ≥ 99%), coumarin (>99% purity), esculin (≥98% purity) were purchased from Sigma-Aldrich (St. Louis, MO, USA), scopoletin (>95% purity) and esculetin (>98% purity) from Extrasynthese (Genay, France), skimmin (98% purity) from Biopurify Phytochemicals (Chengdu, China), scopolin (>98% purity) from Chemicals Aktin Inc. (Chengdu, China). Stock solutions of each standard at a concentration of 10 mmol/L were prepared by diluting the powder in dimethyl sulfoxide (Fisher scientific, Illkirch, France) and kept at −18 °C until use. HPLC-grade methanol was purchased from CarloErba Reagents (Val de Reuil, France), formic acid was purchased from Fisher Scientific (Illkirch, France). Water was purified by a PURELAB Ultra system (Veolia Water S.T.I., Antony, France).
8
Analytical Characterization of Plant Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents used were of at least analytical quality. The standards scopoletin, (+)-catechin, quercetin, kaempferol, taxifolin, eriodictyol, astragalin, isoquercitrin and esculin, as well as methanol, were procured from Merck/Sigma Aldrich (Rahway, NJ, USA). LC-MS grade acetonitrile, water and formic acid were purchased from Fisher Scientific (Geel, Belgium). Vendors for the components of the woody plant medium (WPM), hormones and vitamins are listed in the Supplementary Materials .
9
Phenolic Compounds Dissolution Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The selected phenolic compounds coumarin, esculin and esculetin were purchased from Sigma-Aldrich (Sigma-Aldrich Chemical, Madrid, Spain) and dissolved in DMSO (dimethylsulphoxide) (Dismadel, Madrid, Spain) to a final concentration of 0.5 mM. This concentration is similar to the daily dose clinically used for the flavonoid diosmin (Daflon® 500 mg) [28 (link)] and was established considering the structural similarity (presence of the benzo-α-pyrone core), the almost identical physicochemical properties and their comparable molecular weight between this drug and the compounds here investigated [38 (link)]. Further dilutions were performed to reach 0.75, 1.5, 3.0, 5.0, 6.5 mM for the different assays. To avoid altering the platelet configuration, the lowest volume of DMSO (Dismadel S.L., Madrid, Spain) that could ensure the dissolution of the compounds (2 µL) was added to the blood samples [32 (link)].
10
Melanogenesis Regulation in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Arbutin, N‐phenylthiourea (PTU), tetradecanoyl phorbol acetate (TPA), l ‐DOPA (l ‐3,4‐dihydroxyphenylalanine), l ‐tyrosine, Tyr, sodium hydroxide (NaOH), thiazolyl blue tetrazolium bromide (MTT), Tween‐20, hydrogen chloride (HCl), tricaine methanesulfonate, synthetic melanin, protease inhibitors, TRIS base, dithiothreitol (DTT), bromophenol blue, glycerol, 2‐mercaptoethanol, dimethyl sulfoxide (DMSO), isobutylmethylxanthine (IBMX), rosmarinic acid, rutin hydrate, (‐)quinic acid, baicalein, coumaric acid, epicatechin, ferulic acid, sesamol, protocatechuic acid, quercetin, phloretin and esculin were obtained from Sigma‐Aldrich Co. (St. Louis, MO, USA). Anti‐Tyr (C‐19), anti‐TRP1 (G‐17), anti‐TRP2 (D‐18), anti‐goat IgG‐HRP, anti‐mouse IgG‐HRP or anti‐β‐actin antibody (Santa Cruz Biotech., Santa Cruz, CA, USA) was used. All other reagents and chemicals were high‐grade and commercially available.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!