Silica gel 60 thin layer chromatography plate

Manufactured by Merck Group

Sourced in Germany

Silica gel 60 thin layer chromatography plates are a laboratory equipment used for thin layer chromatography (TLC) analysis. They consist of a uniform layer of silica gel 60 coated on a plastic or aluminum backing, providing a stationary phase for the separation and analysis of chemical compounds.

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6 protocols using silica gel 60 thin layer chromatography plate

In vitro assay for ceramide synthase

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NBD-sphinganine and palmitoyl-CoA (C16-CoA) were from Avanti Polar Lipids (Alabaster, AL). Defatted-bovine serum albumin, a protease inhibitor cocktail and an anti-CerS6 antibody were from Sigma-Aldrich (St. Louis, MO). Horseradish peroxidase was from the Jackson Laboratory (Bar Harbor, ME). An enhanced chemiluminescence (ECL) detection system was from Cyanagen (Bologna, Italy). An anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody and silica gel 60 thin layer chromatography plates were from Merck (Billerica, MA). All solvents were of analytical grade and purchased from Bio-Labs (Jerusalem, Israel).

Kim J., Pewzner-Jung Y., Joseph T., Ben-Dor S, & Futerman A.H. (2022). Generation of a ceramide synthase 6 mouse lacking the DDRSDIE C-terminal motif. PLoS ONE, 17(7), e0271675.

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Lipid Biosynthesis Pathway Probing

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NBD-Sphinganine and fatty acyl CoAs were from Avanti Polar Lipids. Defatted-bovine serum albumin, a protease inhibitor cocktail, anti-HA, anti-CerS2, and anti-tubulin antibodies were from Sigma-Aldrich. Protein A agarose beads and anti-CerS6 antibodies were from Santa Cruz. Horseradish peroxidase was from the Jackson Laboratory. An ECL detection system was from Cyanagen. Silica gel 60 thin layer chromatography plates were from Merck. All solvents were of analytical grade and purchased from Bio-Lab.

Kim J.L., Ben-Dor S., Rosenfeld-Gur E, & Futerman A.H. (2021). A novel C-terminal DxRSDxE motif in ceramide synthases involved in dimer formation. The Journal of Biological Chemistry, 298(2), 101517.

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Thrombin-Induced Lipid Signaling Pathway

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α-Thrombin (human plasma, ≥2,800 NIH units/mg protein) was purchased from Calbiochem (La Jolla, CA, USA) and Sigma-Aldrich (St. Louis, MO, USA). [9,10-³H(N)]palmitic acid, [5,6,7,8,9,11,12,14,15-³H(N)]arachidonic acid was purchased from Amersham Biosciences (Buckinghamshire, England, UK). MTT (Thiazolyl Blue Tetrazolium Bromide), C₆-ceramide, bacterial sphingomyelinase, and 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) were purchased from Sigma-Aldrich. BAPTA-AM (1,2-bis-(o-Aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester) was purchased from BIOMOL Research Labs., Inc (Ann Arbor, MI, USA). Silica gel 60 thin-layer chromatography (TLC) plates were purchased from Merck (Darmstadt, Germany). For the protein assay, the Bradford reagent was purchased from Bio-Rad (Hercules, CA, USA). Plasticware for tissue culture was purchased from NUNC (Waltham, MA, USA). The cleaved caspase-3 ELISA kit was purchased from Cell Signaling Technology (Danvers, MA, USA). AA and Fumonisin B₁ were purchased from Sigma-Aldrich. Trolox was obtained from Tocris (Minneapolis, MN, USA) and the Fluo-4 NW Calcium Assay Kit was purchased from Invitrogen (Carlsbad, CA, USA).

Park M.J., Won J.H, & Kim D.K. (2022). Thrombin Induced Apoptosis through Calcium-Mediated Activation of Cytosolic Phospholipase A2 in Intestinal Myofibroblasts. Biomolecules & Therapeutics, 31(1), 59-67.

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Lipid Extraction and Derivatization

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Phytol, stigmasterol, and β-sitosterol containing campesterol and the fatty acids including linoleic and linolenic acids, myristoyl, palmitoyl, stearoyl, oleoyl chlorides, and N, O-bis (trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane (BSTFA-TMCS, for the trimethylsilyl derivatization) were purchased from Tokyo Chemical Industry Co, Ltd., (TCI). Primuline (for lipid detection) and other organic solvents were purchased from Wako Pure Chemical Industries, Ltd., and silica gel 60 thin-layer chromatography (TLC) plates were purchased from Merck Millipore.

Kondo S., Hori K., Sasaki-Sekimoto Y., Kobayashi A., Kato T., Yuno-Ohta N., Nobusawa T., Ohtaka K., Shimojima M, & Ohta H. (2016). Primitive Extracellular Lipid Components on the Surface of the Charophytic Alga Klebsormidium flaccidum and Their Possible Biosynthetic Pathways as Deduced from the Genome Sequence. Frontiers in Plant Science, 7, 952.

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Reuterin Purification and Standardization

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Three ml of reuterin produced by L. reuteri as described above was taken to prepare the reuterin standard using purification on a silica gel column as previously described^{51 (link)}. The elution was monitored by adding a droplet of the eluent on a silica gel 60 thin-layer chromatography plate (Merck), which was immediately dipped into a 2% (w/v) Purpald reagent solution (Aldrich) in 1 M NaOH. Fractions with reuterin were combined and the solvent was evaporated under reduced pressure. The dry residue was weighed and immediately redissolved in double-distilled water to 2 mg ml⁻¹. The obtained solution was stored at −20 °C until preparation of the standard curve.

Moraïs S., Mazor M., Tovar-Herrera O., Zehavi T., Zorea A., Ifrach M., Bogumil D., Brandis A., Walter J., Elia N., Gur E, & Mizrahi I. (2023). Plasmid-encoded toxin defence mediates mutualistic microbial interactions. Nature Microbiology, 9(1), 108-119.

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Purification of Toxoflavin by TLC

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Toxoflavin was extracted from overnight cultures using chloroform as previously described (Yoneda et al., 1971 ). Chloroform extracts were dissolved in dimethyl sulphoxide and applied to a silica gel 60 thin layer chromatography plate (Merck). Chromatograms were developed with chloroform/methanol (95:5, vol/vol). The spots were visualized under UV light at 365 nm.

Marunga J., Kang Y., Goo E, & Hwang I. (2022). Hierarchical regulation of Burkholderia glumae type III secretion system by GluR response regulator and Lon protease. Molecular Plant Pathology, 23(10), 1461-1471.

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