Silica tlc plate

Manufactured by Merck Group

Sourced in Germany

Silica TLC plate is a thin-layer chromatography (TLC) plate coated with a layer of silica gel. It is used for the separation and identification of compounds in a mixture by means of differential migration.

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Lab products found in correlation

Silica gel, by Merck Group (1 mentions) Imagequant tl software, by GE Healthcare (1 mentions) 1 14c oleoyl coa, by PerkinElmer (1 mentions) Typhoon fla 7000, by GE Healthcare (1 mentions) Phosphorimager, by Fujifilm (1 mentions) 3h serine, by American Radiolabeled Chemicals (1 mentions) Chemidoc, by Bio-Rad (1 mentions)

Close spelling variants of this product

TLC silica gel 60F254 plates Silica gel G-coated TLC plates Silica 60 F254 TLC plates TLC silica gel 60 F 254 aluminum plates TLC silica gel 60 F254 glass plates Silica gel-coated TLC plates TLC silica gel 60 F254 TLC plates Aluminum silica gel 60 F254 TLC plates Precoated silica gel TLC plates TLC silica gel plates 60 F254

12 protocols using silica tlc plate

Lipase Catalysis Assay Protocol

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Example 1

This Example describes a protocol for determining the ability of lipases to catalyze a specific desired reaction. A grid of substrate molecules is prepared in test tubes by dissolving each substrate to a final concentration of 0.1 mM in CH₃CN and mixing with 0.1 M phosphate buffer (pH 7.5). An enzyme is then added to each microtiter well and the mixture is incubated for 30 minutes. The reaction mixture in each tube is extracted three times with dichloromethane, the organic extracts are combined, dried over MgSO₄, and concentrated to about 0.1 mL using a nitrogen stream. The concentrated samples are then analyzed using analytical thin layer chromatography (TLC). Accordingly, each sample is spotted to a silica TLC plate (available from E. Merck, Darmstadt) and developed in a mixture of dichloromethane:methanol (99:1). The developed TLC plates are visualized using UV light and charring with a p-anisaldehyde stain (18 mL p-anisaldehyde, 7.5 mL glacial acetic acid, 25 mL concentrated H₂SO₄, 675 mL absolute ethanol).

US11230722B2. Nutritional supplements and therapeutic compositions comprising (r)-3-hydroxybutyrate derivatives (2022-01-25). Oxford University Innovation Limited [GB], U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT [US]. Inventors: Richard Lewis Veech [US], Michael Todd King [US], Kieran Clarke [GB].

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Thin Layer Chromatography Analysis

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TLC was carried out on a silica TLC plate (Merck, Darmstadt, Germany) in a glass chamber containing organic solvent (as described in the text). The spot on the TLC plate was visualized under UV light or by spraying phosphomolybdic acid.

Cahyono A.W., Fitri L.E., Winarsih S., Prabandari E.E., Waluyo D., Pramisandi A., Chrisnayanti E., Dewi D., Siska E., Nurlaila N., Nugroho N.B., Nozaki T, & Suciati S. (2023). Nornidulin, A New Inhibitor of Plasmodium falciparum Malate: Quinone Oxidoreductase (PfMQO) from Indonesian Aspergillus sp. BioMCC f.T.8501. Pharmaceuticals, 16(2), 268.

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TLC-Direct Bioautography for Antibacterial Screening

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TLC-direct bioautography for antibacterial activity was carried out by the dot-blot method and developed TLCplates aseptically. The antibacterial activity of extracts was carried out against Escherichia coli Ina-CC B5 and Staphylococcus aureus B4. Dot-blot method: Ten µL of extract (10 µg/mL) was transferred onto silica TLC-plate (Merck, silica gel 60 F254) and air-dried. Chloramphenicol was used as a positive control, while the solvent used as the negative control. Developed-TLC plates: The extracts were developed with eluent system as follows: hexane extract was developed with eluent system of hexane: ethyl acetate (3: 1), dichloromethane and ethyl acetate extracts were developed with dichloromethane: methanol (10: 1), while methanol and water extracts were developed with chloroform: methanol: water (6: 4: 1). After finish transferring and developing the extracts, TLC-plate were dipped in the bacterial suspension, and then incubated for 18-24 hours at 37 o C under humid condition by adding sterile wet-cotton. After incubation, plates were sprayed with iodonitrotetrazolium (4 mg/mL) aqueous solution. Growth inhibition was indicated by white zones against the purple background.

Praptiwi P., Wulansari D., Fathoni A., HARNOTO N., Novita R., Alfridsyah A., & Agusta A. (2020). Phytochemical screening, antibacterial and antioxidant assessment of Leuconotis eugenifolia leaf extract. Nusantara Bioscience, 12(1).

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Plant Extract Fractionation by TLC and Column Chromatography

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The thin-layer chromatography (TLC) and column chromatography methods have been widely used to separate fractions from plant extracts. The silica TLC plate and silica gel were purchased from Merck KGaA (Darmstadt, Germany). The TLC method in this research followed the previous study (Kagan and Flythe, 2014) (link). Briefly, the plant extract was dissolved in methanol to have a sample concentration. The TLC plate was cut in a piece (2×10 cm) and leaving a 1 cm border on the upperand bottom sides of the plate. The sample was loaded by using a microliter syringe on the plate band and allow to dry. The plate with the sample was then developed in the cover chamber which containing mixed solvents or eluent (methanol-ethyl acetate-acetone 1:1:1, v/v/v, for this study). After the developing step, the plate was dried and observed bands under visible or UV light (254 nm and 365 nm) then marked bands with a pencil. The mixed solvent was used to further separation by using silica gel column chromatography. Whereas, the column chromatography was performed by the following previous method (Venkatesh et al., 2017) (link). The mixed solvent was loaded into the packed silica column. The seven different separated fractions (confirmed by TLC) were collected, dried, and kept for future analysis.

Herpandı H., Lestari S.D., Bastian, & Sudirman S. (2021). Antioxidant activity of the fractions from water lettuce (Pistia stratiotes) extract. Food Research, 5(2), 451-455.

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ACAT1 Activity Assay by TLC

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An assay in which the activity of human acetyl-coenzyme A acetyltransferase 1 (ACAT1) was detected by thin-layer chromatography (TLC) was performed as described previously. 25 In total, 5 μM [1-14 C]-oleoyl-CoA (PerkinElmer) and 100 μM cholesterol were reacted with 30 μg/mL human ACAT1 enzyme for 40 min at 32 °C. The modified Bligh and Dyer method was applied to the sample preparations. 26 The organic phase was applied to a silica TLC plate (Merck KGaA, Darmstadt, Germany) and separated with a solvent system of hexane, ethyl ether, and acetic acid. The radioactivities incorporated into the lipids were measured with a Typhoon FLA 7000 (GE Healthcare UK Ltd., Little Chalfont, England) and analyzed with Image Quant TL software (GE Healthcare UK Ltd.).

Yoneyama-Hirozane M., Deguchi K., Hirakawa T., Ishii T., Odani T., Matsui J., Nakano Y., Imahashi K., Takakura N., Chisaki I., Takekawa S, & Sakamoto J. (2018). Identification and Characterization of a New Series of Ghrelin O-Acyl Transferase Inhibitors. SLAS discovery : advancing life sciences R & D, 23(2).

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TLC Analysis of ABCA7 Lipid Profiles

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For TLC analysis of ABCA7, lipid extraction was carried out on detergent purified protein by mixing 0.15 mg of protein with chloroform and methanol at a 1:2:2 chloroform:methanol:protein volumetric ratio. The lipids in the lower chloroform phase were collected and dried under argon gas. The lipids were further dissolved in 20 μl of chloroform and 10 μl sample volume was applied to a silica TLC plate (Sigma) together with lipid standards including L‐α‐phosphatidylethanolamine (Brain PE), L‐α‐phosphatidylcholine (Brain PC), L‐α‐phosphatidylserine (Brain PS) (Avanti) that were previously used for nanodisc preparation. The different lipids were separated under a mobile phase (chloroform:methanol:water at 65:25:4 volumetric ratio) and stained with iodine vapor (Sigma) for visualization.

Le L.T., Thompson J.R., Dehghani‐Ghahnaviyeh S., Pant S., Dang P.X., French J.B., Kanikeyo T., Tajkhorshid E, & Alam A. (2022). Cryo‐EM structures of human ABCA7 provide insights into its phospholipid translocation mechanisms. The EMBO Journal, 42(3), e111065.

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Analyzing Phospholipid Binding in the Mla System

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To analyse PL movement within the Mla system a Ni-NTA affinity column protocol coupled with thin layer chromatography (TLC) was employed. To measure whether MlaC could bind PL directly from the environment, without the need for MlaD or MlaFEDB Samples were adjusted to 0.5 mg•mL -1 and 2 mL and added to 2 mL methanol and 1 mL chloroform (2:2:1 v/v). Samples were then vortexed continuously for 5 min, incubated for 30 min at 50°C, and vortexed again for 5 min. The mixture was centrifuged (2000 x g, 10 min), and the lower phase extracted and evaporated. Dried lipids were resuspended in 100 µL chloroform and 5 µL loaded onto a Silica TLC plate (Sigma) and run with a 6.5:2.5:1 (chloroform:methanol:acetic acid) solvent. The TLC plate was dried for 30 min, stained with 10% (w/v) PMA in ethanol, and heated until staining occurred.

Hughes G.W., Hall S.C.L., Laxton C.S., Sridhar P., Mahadi A.H., Hatton C., Piggot T.J., Wotherspoon P.J., Leney A.C., Ward D.G., Jamshad M., Spana V., Cadby I.T., Harding C., Isom G.L., Bryant J.A., Parr R.J., Yakub Y., Jeeves M., Huber D., Henderson I.R., Clifton L.A., Lovering A.L, & Knowles T.J. (2019). Evidence for phospholipid export from the bacterial inner membrane by the Mla ABC transport system. Nature microbiology, 4(10).

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Enzymatic Assay of PI(3,4)P2 Hydrolysis

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Silica TLC plates (Merck Millipore) were preimpregnated in a solution of 2 mM EDTA and 1% potassium oxalate/MeOH (2:3). 48% PC/47.5% PS/4% PI(3,4)P₂/0.5% C16-NBD PI(3,4)P₂ liposomes were prepared to a 2-mM total lipid concentration and extruded through 100-nm or 800-nm polycarbonate filters. The 800-nm extruded population had a mean diameter of 250 nm. 110 µl of each liposome suspension was incubated with 10 nM INPP4A at room temperature in the dark in a buffer containing 150 mM NaCl, 20 mM Hepes, pH 7.4, 2 mM EDTA, and 5 mM CaCl₂. 10-µl samples were taken after 0, 1, 2, 5, 10, 20, 30, 60, 90, 120, and 150 min and immediately boiled to inactivate the enzyme. For each time point sample, lipids were extracted by adding 10 µl chloroform, shaking vigorously, and the organic phase was collected. The extraction was repeated, and the organic phases were pooled, with 10 µl lipid solution in chloroform for each time point spotted on a TLC plate. The plate was eluted with 65% propan-1-ol/35% 2 M acetic acid for several hours in the dark, after which the plates were read on a phosphorimager (Fujifilm) with the band intensity analyzed using the gel tool in ImageJ. Graphs were made in Excel (Microsoft).

Daste F., Walrant A., Holst M.R., Gadsby J.R., Mason J., Lee J.E., Brook D., Mettlen M., Larsson E., Lee S.F., Lundmark R, & Gallop J.L. (2017). Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature. The Journal of Cell Biology, 216(11), 3745-3765.

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Enzymatic Algal Extract Analysis

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Enzymes used on algal extracts are listed in Table S1. All chemical reagents were purchased from Sigma Aldrich, Fisher Scientific, and VWR. All water used was deionised water unless otherwise stated. Silica TLC plates (plastic and aluminium‐backed) were sourced from Merck.

Rapin M.N., Murray L., Sadler I.H., Bothwell J.H, & Fry S.C. (2023). Same but different — pseudo‐pectin in the charophytic alga Chlorokybus atmophyticus. Physiologia Plantarum, 175(6), e14079.

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Measuring Ceramide-to-IPC Conversion in Yeast

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To measure the conversion of ceramide to IPC, cells were labeled with [³H]serine (American Radiolabeled Chemicals, Inc.) as previously described (Kajiwara et al., 2008 (link), 2014 (link)) with the following modifications. In brief, 20 µCi [³H]serine was added to 10 OD₆₀₀ of cultures in mid-logarithmic growth phase in SC without serine at 25°C, and the cells were grown for 1 h. For experiments with strains containing the temperature-sensitive sec18-1 allele, cells were initially grown at 25°C, 200 µg/ml cycloheximide was added to the medium, the cells were grown for 20 more minutes at 25°C, the culture was shifted to 37°C for 30 min, and then labeled for 30 min with 20 µCi [³H]serine (Funato and Riezman, 2001 (link)). After labeling, growth was stopped by addition of 10 mM NaF and 10 mM NaN₃ and placing cultures on ice for 20 min. The cells were washed with water, and lipids were extracted and subjected to mild alkaline hydrolysis to deacylate glycophospholipids as described (Kajiwara et al., 2008 (link), 2014 (link)). Lipids were dried under nitrogen and separated on silica TLC plates (EMD Millipore) using the chloroform/methanol/4.2 N ammonium hydroxide (9:7:2). TLC plates were scanned on a RITA* Thin Layer Analyzer (Raytest) to quantify radiolabeled lipids and determine the ratio of IPC to ceramide.

Liu L.K., Choudhary V., Toulmay A, & Prinz W.A. (2017). An inducible ER–Golgi tether facilitates ceramide transport to alleviate lipotoxicity. The Journal of Cell Biology, 216(1), 131-147.

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