Samples fixed in 4% PFA were washed in 0.1 M PBS for 1 h prior to incubation in 20, 50, 80, and 100% methanol (Merck Millipore, Burlington, MA, United States) in 0.1 M PBS at room temperature for 1 h each. Samples were then washed further with 100% methanol at 4°C for 2 h, then treated with 5% hydrogen peroxide (Merck Millipore, Burlington, MA, United States) at 4°C for 12 h. Samples were subsequently rehydrated in 80, 60, 40, and 20% methanol at room temperature for 1 h each, washed with 0.1 M PBS, and then dehydrated in 20, 40, 60, 80, and 100% methanol at room temperature for 1 h each. Upon dehydration, samples were incubated in 66% DCM:33% methanol at room temperature for 3 h, followed by incubation in 100% DCM for 15 min. Samples were washed with 100% methanol before incubation in DBE for 1–2 days at room temperature until they achieved optical transparency.
Methanol
Manufactured by Merck Group
Sourced in Germany, United States, Italy, India, United Kingdom, China, France, Poland, Spain, Switzerland, Australia, Canada, Sao Tome and Principe, Brazil, Ireland, Japan, Belgium, Portugal, Singapore, Macao, Malaysia, Czechia, Mexico, Indonesia, Chile, Denmark, Sweden, Bulgaria, Netherlands, Finland, Hungary, Austria, Israel, Norway, Egypt, Argentina, Greece, Kenya, Thailand, Pakistan
Methanol is a clear, colorless, and flammable liquid that is widely used in various industrial and laboratory applications. It serves as a solvent, fuel, and chemical intermediate. Methanol has a simple chemical formula of CH3OH and a boiling point of 64.7°C. It is a versatile compound that is widely used in the production of other chemicals, as well as in the fuel industry.
Automatically generated - may contain errors
Lab products found in correlation
Acetonitrile, by Merck Group (2040 mentions)
Formic acid, by Merck Group (1060 mentions)
Ethanol, by Merck Group (1006 mentions)
Chloroform, by Merck Group (819 mentions)
Acetic acid, by Merck Group (628 mentions)
Hydrochloric acid (hcl), by Merck Group (627 mentions)
Sodium hydroxide (naoh), by Merck Group (604 mentions)
Acetone, by Merck Group (561 mentions)
Gallic acid, by Merck Group (510 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (451 mentions)
Ethyl acetate, by Merck Group (430 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (356 mentions)
Hplc grade acetonitrile, by Merck Group (327 mentions)
N hexane, by Merck Group (318 mentions)
Sodium chloride (nacl), by Merck Group (309 mentions)
Sodium carbonate, by Merck Group (306 mentions)
Quercetin, by Merck Group (306 mentions)
Milli q system, by Merck Group (286 mentions)
Dichloromethane, by Merck Group (280 mentions)
Ascorbic acid, by Merck Group (242 mentions)
7 856 protocols using methanol
1
Tissue Clearing and Optical Transparency
2
Extraction and Characterization of Calotropis procera Leaves
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The seeds of C. procera were provided and cultivated in the greenhouse (Fig. 1 ) by Zarringiah Co., West Azerbaijan Province, Urmia, Iran. The growing seedlings were authenticated by a taxonomist. Voucher specimens (voucher numbers: CP/1397 433) were deposited in the Zarringiah Co. Herbarium, Urmia, Iran. The leaves of 6 weeks seedlings were carefully harvested for extraction (Fig. 1 ). After washing and shade drying at room temperature, the samples (500 mg) were powdered and extracted by 10 ml 80% methanol (Merck, Darmstadt, Germany) maceration method and shaking incubation at 25 ± 2 C for 72 h. The extract was paper filtered and the residue re-macerated for the second (80% methanol, 48 h) and third (80% methanol, 24 h) time. Finally, the solvent was evaporated in a vacuum rotary evaporator (Rotary Evaporator N-1110, Eyela, Tokyo, Japan). The concentrated residue was frozen at − 20 C. The dried powders were dissolved in phosphate-buffered saline (PBS, Cl2H3K2Na3O8P2, 1X, pH 7.4, Gibco, Paisley, UK) and diluted to prepare test concentrations of extract.![]()
Seeds and seedlings of Calotropis procera
3
Skin Penetration of Chlorogenic Acid
Check if the same lab product or an alternative is used in the 5 most similar protocols
1% of chlorogenic acid (Sigma-Aldrich, St. Louis, USA) was solubilized in a mixture (2/8, v/v) of PEG-400 (Cooper, Melun, France) and methanol (Sigma-Aldrich, St. Louis, USA). 20 μL of this solution were added to skin samples and incubated in a thermostated Franz cell for 24 hours. The donor compartment of the cell was covered with parafilm to prevent evaporation of the applied compounds. After 24 hours, skin samples were dried; surfaces were gently dabbed with methanol (Sigma Aldrich) using gauze prior to embedding in OTC matrix (CellPath, UK) and frozen in liquid nitrogen. The samples were then stored at −80°C until preparation of microscope slides using a cryostat. Six-micron cryo-cross-sections were observed by fluorescence microscopy (Leica DMR-Camera Leica DFC 310 FX, Nanterre, France) with a DAPI filter (excitation 350 nm and 450–490 nm emission) with few drops of Neu reagent [36 ] (1% of 2-aminoethyl-diphenylborinate) (Sigma-Aldrich, St. Louis, USA) in methanol (Sigma-Aldrich, St. Louis, USA), which potentiates the fluorescence of chlorogenic acid.
+ Expand
1% of chlorogenic acid (Sigma-Aldrich, St. Louis, USA) was solubilized in a mixture (2/8, v/v) of PEG-400 (Cooper, Melun, France) and methanol (Sigma-Aldrich, St. Louis, USA). 20 μL of this solution were added to skin samples and incubated in a thermostated Franz cell for 24 hours. The donor compartment of the cell was covered with parafilm to prevent evaporation of the applied compounds. After 24 hours, skin samples were dried; surfaces were gently dabbed with methanol (Sigma Aldrich) using gauze prior to embedding in OTC matrix (CellPath, UK) and frozen in liquid nitrogen. The samples were then stored at −80°C until preparation of microscope slides using a cryostat. Six-micron cryo-cross-sections were observed by fluorescence microscopy (Leica DMR-Camera Leica DFC 310 FX, Nanterre, France) with a DAPI filter (excitation 350 nm and 450–490 nm emission) with few drops of Neu reagent [36 ] (1% of 2-aminoethyl-diphenylborinate) (Sigma-Aldrich, St. Louis, USA) in methanol (Sigma-Aldrich, St. Louis, USA), which potentiates the fluorescence of chlorogenic acid.
4
RA Metabolism and 4-Hydroxylase Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Overall experimental procedure followed the previous literature in which observed that culture conditions under serum or not, and passage critically affect RA metabolism and induction of 4-hydroxylase activity [19 (link)]. Briefly, HaCaT was seeded and cultured in serum-containing medium for 48 h. Cells at confluency were treated with RA 1 µM (Sigma-Aldrich, St. Louis, MI, USA) and methanol (Sigma-Aldrich, St. Louis, MI, USA) and incubated for additional 48 h. Cultured media and cell lysate were freeze-dried and resuspended in methanol. This methanol-based solution was analyzed by HPLC (Shimadzu, SCL-40, LC-40DXR, CTO-40C etc., Kyoto, Japan). Detailed procedure followed the previous research [20 (link)]. 60 mM ammonium acetate (Sigma-Aldrich, St. Louis, MI, USA) adjusted to pH 5.75 with acetic acid (Sigma-Aldrich, St. Louis, MI, USA) and methanol (Sigma-Aldrich, St. Louis, MI, USA) were used as HPLC buffers. The flow rate was 1 mL/min. The gradient conditions were (1) 15% of methanol at the time of injection, (2) linear increase to 99% of methanol at the 30 min, and (3) maintenance of 99% of methanol for 15 min. UV detection was carried out at 340 nm.
5
Zebrafish Embryo Fixation and In Situ Hybridization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryos were raised at 28°C in petri dishes containing E3 solution. The E3 was changed daily, and any dead embryos removed. At 30–32 hpf, embryos were anaesthetised using tricaine and dechorionated before returned to fresh E3 solution. At the relevant timepoint embryos were fixed overnight at 4°C using 4% (w/v) paraformaldehyde (Sigma-Aldrich, UK) in phosphate buffered saline (PBS). Embryos were washed in PBS/0.05% (v/v) Tween 20 (PBST), then put through a methanol/PBS series using 30%, 60% and 100% (v/v) methanol before being stored in 100% methanol (Sigma-Aldrich) at -20°C. In situ hybridisation (ISH) was carried out as described by [22 (link)], except for the embryo digestion with proteinase K, for which 30–32 hpf embryos were digested with 10 mg/ml proteinase K at 20°C for 22 min. Primers used for PCR generation of the in situ probes are given in Table 1 below. The protocol was conducted with the embryos in 1.5 ml microfuge tubes for the first two days, after which they were held in 12-well plates for staining before transferring back to microfuge tubes for storage. Stained embryos were stored in the dark in 80% (v/v) glycerol.
6
Extraction and Fractionation of E. ivorense
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The leaf and stem bark of E. ivorense were extracted using 99.8% v/v methanol (Sigma-Aldrich, MO, USA). Acetone (96% v/v), petroleum ether (96 v/v), ethyl acetate (98% v/v), and methanol (all solvents purchased from Sigma-Aldrich, MO, USA) were used for the successive extraction and fractionation of the methanol leaf and bark extracts using column chromatography. The extracts were prepared by the cold maceration of 300 g of powdered dry plant material in stoppered flasks containing 700 mL of solvent for 1 week at room temperature (28°C). After filtration, the solvent was evaporated under reduced pressure in a rotary evaporator at 40° C. The different extracts and fractions were conserved in tightly sealed glass vials and stored at 4°C.
7
Colorimetric Determination of Total Flavonoids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total flavonoid content was determined based on the aluminum chloride colorimetric method in which quercetin (Sigma-Aldrich, CAS 117-39-5, ≥95% purity by HPLC) was used as a reference compound [21]. Dissolving 1.0 mg quercetin in 10 mL methanol (Merck) by which the stock solution was made, then by serial dilutions, including using methanol, the standard quercetin solutions were prepared. Briefly, 1 mL of aluminum chloride (Merck) (2% w/v) was added to 1 mL diluted extract or standard quercetin solutions separately, and with methanol, the mixture was made up to 10 mL in quantity. Then, the solution was mixed and incubated for 15 min at room temperature. The absorbance of the mixtures was calibrated at 416 nm with a UV-Vis spectrometer. The measurements were carried out in triplicate. The calibration curve was plotted using standard quercetin. The total flavonoids content was estimated from the calibration plot and expressed as milligram of quercetin equivalent (QE) per gram of dry mass.
8
Comprehensive Metabolite Profiling Protocol
9
Biofluid Metabolomic Sampling and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood (serum and plasma) and urine samples were collected at pre-dose, week 4, and week 8. Blood samples (6 mL) were separately drawn into heparinised tubes and serum separating tubes. The samples were centrifuged (4 °C, 1,800 × g, 8 min), and three plasma and serum aliquots (0.8 mL) were stored in Eppendorf tubes at −70 °C until analysis. Urine samples were collected in 15-mL Falcon tubes and stored at −70 °C until analysis. Frozen samples were thawed at 4 °C, and 50 µL of each sample was diluted with 80% methanol (high-performance liquid chromatography grade, Millipore, Bedford, MA, USA) for plasma and 10% methanol for urine samples. The diluted samples were vortexed for 10 min and centrifuged at 18,341 × g for 20 min at 4 °C. The supernatants were subjected to global metabolomic analysis, and pooled samples were used as quality control samples.
10
Quantitative Analysis of Chlorogenic Acid Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For quantitative purposes primary reference standards were used where possible: 3-caffeoylquinic acid (3CQA), 4-caffeoylquinic acid (4CQA), 5-caffeoylquinic acid (5CQA), 3,4-dicaffeoylquinic acid (3,4-diCQA), 3,5-dicaffeoylquinic acid (3,5-diCQA), and 4,5-dicaffeoylquinic acid (4,5-diCQA) were obtained from PhytoLab GmbH & Co. KG (Vestenbergsgreuth, Germany); caffeine (Ph. Eur. Grade), theobromine, mangiferin trigonelline hydrochloride (analytical grade) methanol and ethanol (HPLC grade) from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Qualitative identification was performed using synthetized standards of FQAs and pCoQAs, not commercially-available. Sodium metabisulfite was obtained from VWR International (Fountenay-sous-Boiscedex, France). All solutions were made with milliQwater system (Millipore, Molsheim, France) and methanol (7:3 v:v).
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!