The BV2 immortalized murine microglial cell line was provided by Dr Il-Whan Choi (Department of Microbiology, College of Medicine, Inje University, Busan, Korea). BV2 microglia were maintained in Dulbecco’s modified Eagle’s medium (DMEM; WelGENE, Inc., Gyeongsan, Korea) containing 10% (v/v) fetal bovine serum (WelGENE, Inc.), L-glutamine (2 mM), penicillin (100 U/ml) and 100 µg/ml streptomycin (WelGENE, Inc.) at 37°C in a humidified atmosphere containing 5% CO2 and 95% air. Isorhamnetin (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) was dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich; Merck KGaA) and was adjusted to final concentrations using complete culture medium. The final DMSO concentration was <0.05% in all experiments (i.e., a non-cytotoxic range). To stimulate cells, the medium was replaced with fresh DMEM and 100 ng/ml LPS (Sigma-Aldrich Chemical Co.) was added in the presence or absence of Isorhamnetin for the indicated time periods.
Isorhamnetin
Manufactured by Merck Group
Sourced in United States, Germany, Spain, Italy, China
Isorhamnetin is a chemical compound used in various laboratory applications. It is a flavonoid found in various plants. Isorhamnetin is utilized for its chemical properties, but its specific core function is not provided here to maintain an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Quercetin, by Merck Group (58 mentions)
Kaempferol, by Merck Group (41 mentions)
Rutin, by Merck Group (31 mentions)
Caffeic acid, by Merck Group (29 mentions)
Luteolin, by Merck Group (26 mentions)
Catechin, by Merck Group (26 mentions)
Chlorogenic acid, by Merck Group (24 mentions)
Gallic acid, by Merck Group (24 mentions)
P coumaric acid, by Merck Group (23 mentions)
Epicatechin, by Merck Group (19 mentions)
Ferulic acid, by Merck Group (18 mentions)
Naringenin, by Merck Group (18 mentions)
Apigenin, by Merck Group (18 mentions)
Formic acid, by Merck Group (17 mentions)
Acetonitrile, by Merck Group (16 mentions)
Myricetin, by Merck Group (16 mentions)
Protocatechuic acid, by Merck Group (14 mentions)
Quercitrin, by Merck Group (13 mentions)
Syringic acid, by Merck Group (12 mentions)
Vanillic acid, by Merck Group (12 mentions)
95 protocols using isorhamnetin
1
Murine Microglial Cell Culture and Isorhamnetin Treatment
2
Dose-Dependent Cytotoxicity of Phytochemicals
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All cell lines were seeded in 96 well plates (Costar) at 5 × 103 cells/well and incubated for 24 h prior to treatment. Cells were treated with afuresertib (Selleckchem), isorhamnetin (Merck), formononetin (Merck), pyrogallol (Merck) and taxifolin (Merck) in a 16-point dose response (n = 6) over 72 h, with concentrations ranging from 1 to 500 μM. Appropriate vehicle (0.1% DMSO) and positive cell death controls (10% DMSO) were used. Viability was assessed by adding 1:10 PrestoBlue (ThermoFisher) over 45 min and measuring the resulting fluorescence at 544/590 nm (excitation/emission) on a FLUOstar Omega microplate reader (BMG LABTECH). Fluorescence intensity was normalized against that of control wells to determine percentage viability of drug-treated cells.
3
Analytical Standards for Phenolic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 14 phenolic acids and flavonoids standards with a purity more than 98% were purchased from Sigma. These included (+)-catechin, epicatechin, gallic acid, ferulic acid, chlorogenic acid, caffeic acid, rutin, spinosin, quercetin, phloridzin, isorhamnetin, luteolin, kaempferol, and jujubosideA. In addition, chromatographic pure methanol, formic acid, and acetonitrile were purchased from Merck.
4
Phenolic Compound Standards for HPLC Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phenolic compound standards: gallic acid, syringic acid, protocatechuic acid, caffeic acid, chlorogenic acid, naringenin, epicatechin, catechin, hesperetin, resveratrol, quercetin, rutin, ellagic acid, myricetin, kaempferol, luteolin, isorhamnetin, epigallocatechin, gallocatechin, gallocatechin gallate, epicatechin gallate, procyanidin A1, procyanidin A2, procyanidin B1, procyanidin B2, procyanidin C1, and corilagin were HPLC grade and purchased from Merck (St. Louis, MO, USA). Acetonitrile, formic acid, methanol, hexane, and water were HPLC/spectrum grade and purchased from Tedia (Fairfield, OH, USA). All other reagents and solvents used were analytical grade from Merck (St. Louis, MO, USA).
5
Characterization of Urban Particulate Matter
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quercetin, penitrem A, and isorhamnetin were purchased from MERCK (Darmstadt, Germany). Particulate matter PM < 4 μm SRM-2786 (PM) was obtained from Nation Institute of Standards and Technology (NIST, Gaithersburg, MD, USA). In our study, we used PM according to NIST standard in order to ensure the repeatability of biophysical and biochemical experiments. PM was formulated from atmospheric particulate material gathered in 2005 from an air intake filtration system of a major exhibition center in Prague, Czech Republic. As the producer declares, the PM sample is not expected to represent the area from which it was collected, it should rather reflect atmospheric particulate matter present in an urban area.
6
SH-SY5Y Neuronal Differentiation and Treatments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SH-SY5Y cells (American Type Culture Collection (ATCC) CRL-2266) were maintained in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) containing 10% fetal bovine serum (FBS) at 37°C under 5% CO2. Subsequently, for neuronal differentiation, 20 μM retinoic acid (RA) was added to the culture medium and the cells were incubated for 5 days. The differentiated cells were treated with isorhamnetin (10 μM, Merck), TYK2 inhibitor (1 μM, BMS-986165, AdooQ), or IL-6-neutralized antibody (IL-6 IgG, 5 ng/mL, InvivoGen) for 2 days, respectively.
7
Antioxidant Analysis of Phenolic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All the solvents and standards were of analytical grade. Standards of 3,5-di-caffeoylquinic acid (3,5-diCQA), ferulic acid, 3,5-dimethoxy-4-hydroxycinnamic acid (sinapinic acid), p-coumaric acid, hesperidin, isorhamnetin, catechin, rutin, quercetin were purchased from Sigma-Aldrich (Saint Louis, MO, USA). All the stock solutions of standards, were prepared in methanol and were stored at -20 °C. Further dilutions were prepared using methanol. Chlorogenic acid, Folin-Ciocialteu reagent, ABTS •+ (2, 2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) and Trolox ((±)-6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid) were obtained from Sigma-Aldrich (Saint Louis, MO, USA), acetonitrile from Merck (Darmstadt, Germany), while methanol, acetone, hydrochloric acid, sodium carbonate and formic acid from POCh (Katowice, Poland).
8
Comprehensive Chemical Compound Procurement
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Acetonitrile, dichloromethane and acetone were obtained from VWR (Leuven, Belgium). Methanol, acetic acid, hydrochloric acid, sodium tetraborate, sodium hydroxide and sodium hydrogen carbonate were from Merck (Darmstadt, Germany). Sodium borohydride, N-methyl imidazole, potassium hydroxide, 3-phenylphenol, lignin, acetic anhydride, acetyl bromide, perchloric acid, chlorogenic acid, (þ)-catechin, (À)-epicatechin, quercetin, isorhamnetin, hydroxylamine hydrochloride and benzyl mercaptan were provided by Sigma Aldrich (Steinheim, Germany). Sugar standards were from Fluka-Biochemica (Sigma Aldrich, Steinheim, Germany).
9
Polyphenol Extraction and LC-MS/MS Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The solvents used for polyphenol extraction and liquid chromatographic mass spectrometry (LC-MS/MS) analysis, methanol, acetonitrile and formic acid, were purchased from Merck (Darmstadt, Germany). 2,2-diphenyl-1picrylhydrazyl (DPPH), used to determine the antioxidant capacity, was purchased from Thermo Fisher (Kandel, Germany). The polyphenols used as standards, aminobenzoic acid, acetylsalicylic acid, caffeic acid, chlorogenic acid, ellagic acid, gallic acid, p-coumaric acid, protocatechuic acid, salicylic acid, trans-ferulic acid, vanillic acid, apigenin, epicatechin, aesculetin, catechin hydrate, isorhamnetin, kaempferol, luteolin, polydatin, quercetin, resveratrol, rutin, syringaldehyde and viniferin, were purchased from Sigma-Aldrich (Madrid, Spain). The Mili-Q water used in all the solutions was purified with the Merck Millipore Milli-Q™ Reference Ultrapure Water Purification System model Z00QSVC01 (Darmstadt, Germany).
10
Analytical Characterization of Natural Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analytical-grade methanol and n-hexane for extractions were obtained from Sigma-Aldrich (Milan, Italy). Ultrapure water (18 MΩ) was prepared by a Milli-Q purification system (Millipore, Bedford, MA, USA). methanol (MeOH), water, and formic acid with LC-MS grade were supplied by Romil (Cambridge, UK). Reference standards (>98% HPLC grade) apigenin, luteolin, rutin, coumarin, eriocitrin, isorhamnetin, hesperetin, naringenin, 1,1-diphenyl-2-picrylhydrazyl (DPPH•), and 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS•+) reagents were purchased from Sigma-Aldrich (Milan, Italy). Standard stock solutions (1 mg mL−1) of each compound were prepared in methanol and stored at 4 °C. Diluted solutions and standard mixtures were prepared in MeOH/H2O·2:8 (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!