Tlc plate

Manufactured by Macherey-Nagel

Sourced in Germany

TLC plates are a type of chromatographic media used for thin-layer chromatography (TLC) analysis. They are coated with a thin layer of adsorbent material, typically silica gel or alumina, and are used for the separation and identification of various chemical compounds.

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Lab products found in correlation

Prism 9, by GraphPad (2 mentions) Expressionl cms mass spectrometer, by Advion (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Mevinolin, by Merck Group (1 mentions) Atorvastatin, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Hepg2, by American Type Culture Collection (1 mentions) 1 14c oleic acid, by Cytiva (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Camphene, by Merck Group (1 mentions) Acetohydroxamic acid, by Merck Group (1 mentions) 2 3 dihydroxybenzoic acid, by Merck Group (1 mentions) Cbn d3, by Cerilliant (1 mentions) Triethylamine, by Thermo Fisher Scientific (1 mentions) Formic acid, by Avantor (1 mentions) Cbd d3, by Cerilliant (1 mentions) Chromabond c18 ec, by Macherey-Nagel (1 mentions) Xtra sil g uv254, by Macherey-Nagel (1 mentions) N heptane, by Avantor (1 mentions)

12 protocols using tlc plate

Synthesis and Characterization of Copper(II) Complex

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[Cu(tmpa)(MeCN)](OTf)₂ was afforded
as reported.^{28 (link)} 4-((^tButyldimethylsilyl)oxy)butan-1-ol was obtained from Santa
Cruz Biotechnology and used as received. All other chemicals and solvents
were purchased from Merck or VWR and were used as received as well.
Deoxygenated and anhydrous solvents were obtained from a PureSolv
PS-MD-5 solvent dispenser (Innovative Technology). Column chromatography
was performed on alumina (Al₂O₃, activated,
basic, Brockmann I, 58 Å pore size, pH 9.5 ± 0.5) . Thin
layer chromatography (TLC) was performed using TLC plates from Machery-Nagel
(Alugram Aloz, Al₂O₃, with F₂₅₄ indicator
on aluminum backing, pH 9). Compounds were visualized on TLC plates
by UV detection at 254 nm. ¹H, COSY, ¹³C APT,
HSQC, and HMBC NMR spectra were recorded on a Bruker 400 MHz (100.6
MHz for ¹³C) NMR spectrometer using the residual solvent
as internal standard. Mass spectra were obtained by high resolution
mass spectrometry (HRMS) using a TOF Synapt G2-Si mass spectrometer
equipped with an electrospray ionization source in positive ion mode
with leu-enkephalin (m/z = 556.2771)
as an internal lock mass. TLC/mass spectrometry (TLC/MS) analysis
was performed on an Advion Plate Express TLC Plate Reader connected
to an Advion expression^L CMS mass spectrometer.

Smits N.W., den Boer D., Wu L., Hofmann J.P, & Hetterscheid D.G. (2019). Elucidation of the Structure of a Thiol Functionalized Cu-tmpa Complex Anchored to Gold via a Self-Assembled Monolayer. Inorganic Chemistry, 58(19), 13007-13019.

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Cholesterol Biosynthesis Inhibition Assay

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Camphene, mevinolin, atorvastatin and U18666A were from Sigma-Aldrich. [2-¹⁴C] acetic acid sodium salt (250 μCi) and [1-¹⁴C] oleic acid (50 μCi) were purchased from Amersham Biosciences. ACAT inhibitor, F1394, was kindly provided by Dr Constantinos G. Panousis. Reagents for protein determination were from Bio-Rad. The human hepatic cell line, HepG2, was purchased from ATCC. Cell culture medium, Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and antibiotics, penicillin and streptomycin, were from Gibco BRL Life Technologies. Lipoprotein-deficient serum (LPDS) was obtained from Autogen Bioclear UK Ltd. TLC plates were from Macherey Nagel.

Vallianou I, & Hadzopoulou-Cladaras M. (2016). Camphene, a Plant Derived Monoterpene, Exerts Its Hypolipidemic Action by Affecting SREBP-1 and MTP Expression. PLoS ONE, 11(1), e0147117.

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Thin Layer Chromatography of Plant Extracts

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Extracted plant material (10 μL) dissolved in acetone (10 mg/mL) was spotted onto TLC plates (Macherey-Nagel, Düren, Germany) and eluted in different solvent systems, namely, EMW (ethyl acetate : methanol : water, 10 : 1.35 : 1 v/v/v), BEA (benzene : ethanol : ammonia, 18 : 2 : 0.2, v/v/v), and CEF (chloroform : ethyl acetate : formic acid, 10 : 8 : 2 v/v/v). The plates were sprayed with vanillin-sulphuric acid reagent (0.1 g vanillin, 28 mL methanol, and 1 mL sulphuric acid) and developed in an oven at 110°C for 5 min [10 (link)].

Pilane M.C., Bagla V.P., Mokgotho M.P., Mbazima V., Matsebatlela T.M., Ncube I, & Mampuru L. (2015). Free Radical Scavenging Activity: Antiproliferative and Proteomics Analyses of the Differential Expression of Apoptotic Proteins in MCF-7 Cells Treated with Acetone Leaf Extract of Diospyros lycioides (Ebenaceae). Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 534808.

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Extraction and Identification of Siderophores

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SlFG3 was grown in 10 ml of M9 medium for 12 hours at 28 ± 2 °C. The cells were removed by centrifugation (8000 × g, 10 min). Siderophores were extracted from the supernatant with 10 ml of ethyl acetate, dried and resuspended in 10 μl of 50% aqueous ethanol. Siderophores were then separated on TLC plates (Macherey-Nagel, Düren, Germany), coated with a 0.25 mm layer of silica gel, with methanol/acetonitrile (7:3, v/v). Iron-binding compounds were identified by sputtering the TLC plates with 0.05% ferric chloride in ethanol. The commercial siderophores 2,3-dihydroxybenzoic acid and acetohydroxamic acid (Sigma, Darmstadt, Germany) were used as controls for migration during TLC.

Caneschi W.L., Sanchez A.B., Felestrino É.B., Lemes C.G., Cordeiro I.F., Fonseca N.P., Villa M.M., Vieira I.T., Moraes L.Â., Assis R.D., do Carmo F.F., Kamino L.H., Silva R.S., Ferro J.A., Ferro M.I., Ferreira R.M., Santos V.L., Silva U.D., Almeida N.F., Varani A.D., Garcia C.C., Setubal J.C, & Moreira L.M. (2019). Serratia liquefaciens FG3 isolated from a metallophyte plant sheds light on the evolution and mechanisms of adaptive traits in extreme environments. Scientific Reports, 9, 18006.

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BCDX2 ATPase Activity Assay

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Wild-type or mutant BCDX2 (1 μM) were mixed with 15 μM cold ATP and 0.1 μCi/μL α-^{32 (link)}P-ATP, with and without dN^15nt ssDNA (3 μM) in 25 mM HEPES pH 7.5, 5% glycerol, 100 mM NaCl, 2.5 mM MgCl₂ and 0.25 mM TCEP. Samples were incubated (37°C, 30 min) before quenching with 10 mM EDTA. α-^{32 (link)}P-ATP and α-^{32 (link)}P-ADP were separated by chromatography on TLC plates (Macherey-Nagel) using 500 mM LiCl and 1 M formic acid buffer. Plates were dried, exposed to a phosphoscreen and imaged on a Typhoon 9500 phosphorimager. Percentage ATP hydrolysis was calculated using Fiji (ImageJ). Statistical analyses and figure plotting were performed using GraphPad Prism 9.

Greenhough L.A., Liang C.C., Belan O., Kunzelmann S., Maslen S., Rodrigo-Brenni M.C., Anand R., Skehel M., Boulton S.J, & West S.C. (2023). Structure and function of the RAD51B-RAD51C-RAD51D-XRCC2 tumour suppressor. Nature, 619(7970), 650-657.

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Cannabinoid Standardization and Analysis

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CBN (1 mg/mL in methanol, certified reference material), CBN-d₃ (100 μg/mL in methanol, certified reference material), CBD (1 mg/mL in methanol, certified reference material), and CBD-d₃ (100 μg/mL in methanol, certified reference material) were bought from Cerilliant; acetonitrile (HPLC gradient grade), methanol (HPLC gradient grade), n-hexane (HPLC grade), n-heptane (HPLC grade), and formic acid (98–100%) were obtained from VWR Chemicals; dichloromethane (HPLC grade) from Carl Roth, triethylamine from ACROS Organics, Fast Blue Salt B (FBS, dye content ~ 95%) from Sigma-Aldrich, Chromabond SiOH (1 ml/100 mg), Chromabond C₁₈ ec (1 ml/ 100 mg) as well as TLC (thin-layer chromatography) plates (silica gel 60, ALUGRAM Xtra SIL G UV₂₅₄ and octadecyl-modified silica, ALUGRAM RP-18 W/UV₂₅₄) from Macherey-Nagel, TLC plates (silica gel 60 without fluorescent indicator on aluminum sheets), and HPTLC (high-performance thin-layer chromatography) plates (silica gel 60 F₂₅₄ MS-grade for matrix-assisted laser desorption/ionization (MALDI) and silica gel 60 F₂₅₄ on glass plates) were purchased from Merck KGaA and analytical sea sand from Grüssig GmbH. Distilled diethyl ether and acetone were produced with a rotary evaporator from BÜCHI.

Schmidt T., Kramell A.E., Oehler F., Kluge R., Demske D., Tarasov P.E, & Csuk R. (2020). Identification and quantification of cannabinol as a biomarker for local hemp retting in an ancient sedimentary record by HPTLC-ESI-MS. Analytical and Bioanalytical Chemistry, 412(11), 2633-2644.

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Quantitative Steroid Metabolism Assay

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Steroid metabolism was labeled by adding either 54‘000 cpm [³H]-pregnenolone, 49,000 cpm [³H]-DHEA or 59,000 cpm [¹⁴C]-progesterone per well for 120, 90 and 60 minutes, respectively. Steroids were extracted from medium as previously described [12 (link)] and separated on thin-layer chromatography (TLC) plates (Macherey-Nagel, Düren, Germany) using the chloroform:ethylacetate (3:1) solvent system. The separated steroids were visualized by exposing TLC plates on imaging screens and reading them on a Fuji PhosphoImager FLA-7000 (Fujifilm, Dielsdorf, Germany). Steroids were identified by running known standards in parallel on screens. Specific steroids were densitometrically quantified with the Multi Gauge software (Fujifilm). Finally, specific steroid conversion was calculated as a percentage of radioactivity incorporated in a specific steroid hot spot when compared with the total radioactivity added to the reaction.

Marti N., Bouchoucha N., Sauter K.S, & Flück C.E. (2017). Resveratrol inhibits androgen production of human adrenocortical H295R cells by lowering CYP17 and CYP21 expression and activities. PLoS ONE, 12(3), e0174224.

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Quantifying 11β-HSD2 Activity in JEG-3 Cells

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JEG-3 cells were seeded (50,000 cells/well in 100 µL) into 96-well plates and treated for 24 h.
Subsequently, the medium was replaced by 50 µL charcoal-treated serum-free medium and cells were incubated for 4 h with 50 nM cortisol (containing 10 nCi of [1,2,6,7-3 H]-cortisol). Reactions were J o u r n a l P r e -p r o o f stopped by adding an excess cortisol and cortisone (1:1, 2 mM each in methanol). Supernatant (20 µL) was loaded on TLC plates (Macherey-Nagel, Oensingen, Switzerland) and glucocorticoids were separated using chloroform and methanol (9:1). Liquid scintillation counting was performed to determine 11β-HSD2 dehydrogenase activity and compared to control samples.

Inderbinen S.G., Engeli R.T., Rohrer S.R., Di Renzo E., Aengenheister L., Buerki-Thurnherr T, & Odermatt A. (2020). Tributyltin and triphenyltin induce 11β-hydroxysteroid dehydrogenase 2 expression and activity through activation of retinoid X receptor α. Toxicology letters, 322.

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BCDX2 ATPase Activity Assay

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Sphingomyelin Quantification in Ischemic Brain

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Tissue samples obtained from the reperfused ischemic middle cerebral artery territory and homologous contralateral brain tissue were lysed in 250 mM sodium acetate buffer (pH 5.0) containing 1% NP-40 detergent (Fluka BioChemika, Morristown, NJ, U.S.A.). Cellular membrane integrity was disrupted with a sonicator. After centrifugation for 5 min at 300 g at 4 °C, supernatants were collected. Lysates were adjusted to a specific protein concentration and incubated with 100 pmol BODIPY-labeled sphingomyelin (Thermo Fisher Scientific) in 250 mM sodium acetate (pH 5.0) and 0.1% NP-40 for 1 h at 37 °C. Chloroform:methanol (2:1, v/v) was added, samples were vortexed and centrifuged for 5 min at 15,000 g to achieve a phase separation. The lower phase was collected and concentrated in a vacuum centrifuge (SPC111V, Thermo Fisher Scientific) for 45 min at 37 °C. Lipids were dissolved in 20 µl chloroform:methanol (2:1, v/v) and spotted onto thin layer chromatography (TLC) plates (Macherey Nagel, Düren, Germany). The TLC run was performed with chloroform:methanol (80:20, v/v). TLC plates were analyzed with a Typhoon FLA 9500 scanner (GE Healthcare Life Sciences) and lipid spots were quantified with ImageQuant (GE Healthcare Life Sciences).

Hagemann N., Mohamud Yusuf A., Martiny C., Zhang X., Kleinschnitz C., Gunzer M., Kolesnick R., Gulbins E, & Hermann D.M. (2020). Homozygous Smpd1 deficiency aggravates brain ischemia/ reperfusion injury by mechanisms involving polymorphonuclear neutrophils, whereas heterozygous Smpd1 deficiency protects against mild focal cerebral ischemia. Basic Research in Cardiology, 115(6), 64.

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