The pNP derivatives butyrate, octanoate, decanoate, dodecanoate and tetradecanoate, hexaoctanoate, 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS), cellosolve and fluorescein dilaurate were from Sigma Aldrich (St. Louis, MO, USA). Fluorescein dihexanoate and dioctanoate were synthesized by GalChimia, S.L., (O Pino, A Coruña, Spain, https://www.galchimia.com ). Molecular weight markers for electrophoresis were obtained from Bio-Rad (Richmond, CA, USA). All other chemicals were of the purest grade available.
Octanoate
Manufactured by Merck Group
Sourced in United States
Octanoate is a chemical compound used in laboratory settings. It is a carboxylic acid with the formula CH3(CH2)6COO-. Octanoate serves as a component in various laboratory procedures and experiments, but a detailed description of its core function is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Butyrate, by Merck Group (6 mentions)
Decanoate, by Merck Group (3 mentions)
Hexanoate, by Merck Group (3 mentions)
Palmitate, by Merck Group (2 mentions)
P nitrophenyl acyl esters, by Merck Group (2 mentions)
P nitrophenyl hexanoate, by Tokyo Chemical Industry (2 mentions)
Propionate, by Merck Group (2 mentions)
Cellosolve, by Merck Group (1 mentions)
3 3 cholamidopropyl dimethylammonio 1 propanesulphonate chaps, by Merck Group (1 mentions)
Methyl tertiary butyl ether, by Merck Group (1 mentions)
Tris hcl, by Merck Group (1 mentions)
P nitrophenol, by Merck Group (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by GoldBio (1 mentions)
Polymyxin b sulfate, by Thermo Fisher Scientific (1 mentions)
Kanamycin, by GoldBio (1 mentions)
Granulated agar, by Merck Group (1 mentions)
Coenzyme a, by Merck Group (1 mentions)
Valerate, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
9 protocols using octanoate
1
Synthesis and Characterization of Lipid Derivatives
2
Enzymatic Assay of Acyl CoA Hydrolase
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The following p-nitrophenyl acyl esters were obtained from Sigma-Aldrich (St. Louis, MO): acetate, butyrate, octanoate, decanoate, myristate, and palmitate. p-Nitrophenyl hexanoate was obtained from Tokyo Chemical Industry. Coenzyme A, Tris-HCl, p-nitrophenol, lysogeny broth mix, granulated agar, and methyl tertiary-butyl ether were also obtained from Sigma-Aldrich. Polymyxin B sulfate was obtained from Alfa Aesar. Isopropyl-β-d -1-thiogalactopyranoside (IPTG) and kanamycin were obtained from GoldBio.
3
Maximizing Bioplastic Production in Methylocystis parvus
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Fifty-milliliter Methylocystis parvus OBBP cultures were grown to final optical densities (OD600) of 0.8–1.2 then centrifuged (3000g) for 15 min. The pellets were resuspended in 30 mL of JM2 medium to create the inoculum for triplicate 160 mL serum bottle cultures. Each culture received 10 mL inoculum plus 40 mL of fresh medium (39.5 mL of medium JM2 plus 0.5 mL of 1.35 M ammonium chloride stock) and was flushed for 5 min with a CH4/O2 mixture (molar ratio of 1:1.5). After growth at 30 °C for 24 h, the headspace in each culture was again flushed for 5 min with the CH4/O2 mixture then incubated at 30 °C for a second 24 h period of balanced growth.
After 48 h, all cultures were harvested and subjected to nitrogen-limiting conditions. Triplicate samples were centrifuged (3000g) for 15 min and suspended in fresh medium without nitrogen. The headspace of each bottle was flushed with the CH4:O2 gas mixture at t = 0 h and t = 24 h. To assess the effects of co-substrate addition of PHA synthesis, the medium was amended with varying concentrations of 4HB, 5HV, and 6HHx monomers. Other organic acid co-substrates including 3HB, butyrate, valerate, hexanoate, and octanoate (Sigma-Aldrich, St Louis, MO, USA) were also tested for PHA copolymer synthesis. After 48 h of incubation, cells were harvested by centrifugation (3000g) and freeze-dried. Preserved samples were assayed for PHA content.
After 48 h, all cultures were harvested and subjected to nitrogen-limiting conditions. Triplicate samples were centrifuged (3000g) for 15 min and suspended in fresh medium without nitrogen. The headspace of each bottle was flushed with the CH4:O2 gas mixture at t = 0 h and t = 24 h. To assess the effects of co-substrate addition of PHA synthesis, the medium was amended with varying concentrations of 4HB, 5HV, and 6HHx monomers. Other organic acid co-substrates including 3HB, butyrate, valerate, hexanoate, and octanoate (Sigma-Aldrich, St Louis, MO, USA) were also tested for PHA copolymer synthesis. After 48 h of incubation, cells were harvested by centrifugation (3000g) and freeze-dried. Preserved samples were assayed for PHA content.
4
Culturing HepG2 and Upcyte Hepatocytes
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HepG2 cells were routinely grown in culture grade flasks at 37 °C under a humidified atmosphere 5% CO2/95% air in Ham’s F-12/Leibovitz L-15 (1:1, v/v) supplemented with 7% fetal bovine serum (Capricorn Scientific GmbH, Ebsdorfergrund, Germany), 50 U/mL penicillin (Gibco, Waltham, MA, USA), and 50 μg/mL streptomycin (Gibco). Cells were used or passaged at 70–80% confluence. For subculturing purposes, cells were detached with 0.25% trypsin/0.02% EDTA in PBS at 37 °C.
Human Upcyte® hepatocytes (Upcyte® Technologies GmbH, Hamburg, Germany) are cells derived from primary human hepatocytes by a novel technology that forces cells to a limited proliferation, without immortalization, while maintaining specific liver functions. Unlike HepG2 cells, they express characteristic hepatic genes [5 (link)] and preserve a phenotype close to that of primary cultured human hepatocytes [35 (link),36 (link)]. Upcyte hepatocytes were cultured essentially as previously described [36 (link)].
Cells were treated with the appropriate compound or with an equal volume of vehicle (PBS) for 24 h. Valproate (Cat. no. P4543), butyrate (Cat. no. B5887), octanoate (Cat. no. C5038), hexanoate (Cat. no. C4026), propionate (Cat. no. P1880), and acetate (Cat. no. S2889) were purchase from Sigma-Aldrich (Taufkirchen, Germany).
Human Upcyte® hepatocytes (Upcyte® Technologies GmbH, Hamburg, Germany) are cells derived from primary human hepatocytes by a novel technology that forces cells to a limited proliferation, without immortalization, while maintaining specific liver functions. Unlike HepG2 cells, they express characteristic hepatic genes [5 (link)] and preserve a phenotype close to that of primary cultured human hepatocytes [35 (link),36 (link)]. Upcyte hepatocytes were cultured essentially as previously described [36 (link)].
Cells were treated with the appropriate compound or with an equal volume of vehicle (PBS) for 24 h. Valproate (Cat. no. P4543), butyrate (Cat. no. B5887), octanoate (Cat. no. C5038), hexanoate (Cat. no. C4026), propionate (Cat. no. P1880), and acetate (Cat. no. S2889) were purchase from Sigma-Aldrich (Taufkirchen, Germany).
5
Enzymatic Assays with Acyl Esters
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The p-nitrophenyl acyl esters (acetate, butyrate, octanoate, decanoate, myristate, and palmitate) were obtained from Sigma-Aldrich (Yongin, Korea). All enzymes, including Taq polymerase, thrombin, and gel filtration calibration kits for size exclusion chromatography, were obtained from Sigma-Aldrich (Yongin, Korea). Isopropyl β-d-1-thiogalactopyranoside (IPTG) and kanamycin were purchased from Fisher Scientific (Seoul, Korea).
6
Intestinal Cell Culture with Fatty Acids
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The method of obtaining intestinal cells was similar to Fang et al. (24 (link)). Briefly, 15% fetal bovine serum (FBS) (Biological Industries, Israel) was added into Dulbecco’s modified Eagle’s medium (DMEM)/F12 (Biological Industries, Israel) to culture intestinal cells isolated from healthy large yellow croakers. Cells were cultivated in 27°C and 5% CO2 atmosphere. Intestinal cells were seeded in six-well plates, and after 48 h of culture, intestinal cells were serum-starved with FBS-free DMEM/F12 for 2 h. When intestinal cells completed hungry, different concentrations of docosahexaenoic acid (DHA) (Sigma, USA), linoleic acid (LA) (Sigma, USA), and octanoate (Sigma, USA) were supplied in DMEM/F12 individually or together to culture intestinal cells to simulate the situation in vivo. Then, cells were harvested for further analysis.
7
Adipocyte Differentiation and miR-33b Regulation
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PSPA were cultured in DMEM growth medium containing a low glucose concentration (1.0 g/L) and 10 % fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA, USA). The subconfluent cells were passed every 3 days. To produce mature adipocytes, we plated the PSPA at 2.1 × 104 cells/cm2 and grew them for 3 days to reach confluence. After the cells reached confluence (denoted as 0 day), adipose conversion was carried out in high-glucose (4.5 g/L) DMEM containing 10 % FBS, 5 μg/mL insulin (Sigma-Aldrich, Basel, Switzerland), 0.25 μM dexamethasone (Sigma-Aldrich), 33 μM biotin (Wako Pure Chemicals, Osaka, Japan), 17 μM pantothenate (Sigma-Aldrich), and 5 mM octanoate (Sigma-Aldrich). The medium was changed every other day. To investigate the effect of miR-33b on these cells, we subjected groups of cells to the following three experimental conditions: (1) growth medium alone; (2) differentiation with a negative control miRNA (miR-NC) transfection; and (3) differentiation with miR-33b transfection. All cells were cultured at 37 °C in a humidified incubator in 5 % CO2.
8
Vibrational Spectroscopy of Sodium Carboxylates
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We used sodium carboxylates as received:
formate (≥99.0%, Fluka Analytical), acetate (anhydrous, for
molecular biology ≥99%, Sigma-Aldrich), propionate (minimum
99%, Sigma), hexanoate (99–100%, Sigma), octanoate (≥99%,
Sigma), benzoate (>99%, Sigma-Aldrich), and 2-naphthoate (>98%,
TCI).
To prepare the samples, we dissolved appropriate amounts of the salts
in D2O (99.9% D atom, Aldrich). For each carboxylate, we
chose the concentration to obtain a signal-to-noise ratio that is
more than sufficient to reliably determine the amplitudes of the VSFG
responses of the νs and νas vibrations
of the carboxylate groups in different VSFG polarization combinations.
The concentrations of the solutes are reported in the units of molality
(mol/kg solvent (m)).
formate (≥99.0%, Fluka Analytical), acetate (anhydrous, for
molecular biology ≥99%, Sigma-Aldrich), propionate (minimum
99%, Sigma), hexanoate (99–100%, Sigma), octanoate (≥99%,
Sigma), benzoate (>99%, Sigma-Aldrich), and 2-naphthoate (>98%,
TCI).
To prepare the samples, we dissolved appropriate amounts of the salts
in D2O (99.9% D atom, Aldrich). For each carboxylate, we
chose the concentration to obtain a signal-to-noise ratio that is
more than sufficient to reliably determine the amplitudes of the VSFG
responses of the νs and νas vibrations
of the carboxylate groups in different VSFG polarization combinations.
The concentrations of the solutes are reported in the units of molality
(mol/kg solvent (m)).
9
Biopolymer Synthesis: Reagents & Protocols
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All chemicals used in this study were of analytical grade. Chloroform, acetonitrile, poly-3-hydroxybutyrate-co-3-hydroxy-valerate(P(3HB-co-3HV)) pellets, p-nitrophenyl acetate, butyrate, octanoate, decanoate, and dodecanoate were obtained (Sigma-Aldrich, St. Louis, MO, USA). We obtained p-nitrophenyl hexanoate (Tokyo Chemical Industry, Tokyo, Japan). PHB pellets were obtained(Goodfellow Cambridge Ltd., Huntingdon, UK). PBAT and PBS pellets were obtained (Gio Soltech Co., Ltd. Wonju, Korea). Poly-3-hydroxybutyrate-co-4-hydroxybutyrate (P(3HB-co-4HB)) pellets were obtained (CJ, Suwon, Korea). Dichloromethane (DCM), fructose, and sucrose were obtained (Junsei Chemical Co., Tokyo, Japan). Glucose and xylose were obtained (Duksan Pure Chemicals, Ansan, Korea). Galactose was obtained (Daejung Chemicals, Siheung, Korea).
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