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Celite 545

Manufactured by Thermo Fisher Scientific
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Celite 545 is a diatomaceous earth filter aid product. It is used as a filtration medium to remove particulates from liquids in various industrial applications.

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Lab products found in correlation

Ethyl acetate, by Thermo Fisher Scientific (4 mentions) Acetone, by Thermo Fisher Scientific (4 mentions) Silicic acid, by Merck Group (4 mentions) Calcium phosphate cahpo4.2h2o, by Thermo Fisher Scientific (4 mentions) Bis trimethylsilyl trifluoroacetamide bstfa 1 trimethylchlorosilane tmcs, by Merck Group (4 mentions) Oleic acid, by Merck Group (3 mentions) Linoleic acid, by Merck Group (3 mentions) Methanol, by Thermo Fisher Scientific (3 mentions) Hplc grade hexane, by Thermo Fisher Scientific (3 mentions) Chloroform, by Thermo Fisher Scientific (3 mentions) 7β hydroxycholesterol, by Merck Group (2 mentions) 5 6α epoxycholesterol, by Merck Group (2 mentions) 5 6β epoxycholesterol, by Merck Group (2 mentions) Cholestanetriol triol, by Merck Group (2 mentions) 25 hydroxycholesterol, by Merck Group (2 mentions) 25 gram solid phase cartridge, by Teledyne (2 mentions) Celite, by Teledyne (2 mentions) Hexane, by Thermo Fisher Scientific (1 mentions) Calcium hydride, by Merck Group (1 mentions) Anhydrous sodium sulphate, by Thermo Fisher Scientific (1 mentions)

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Celite®

10 protocols using celite 545

1

Fractionation and purification of bioactive compound

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The fractionation scheme is provided in Figure S1. A portion of the methanol extract (900 mg) was dissolved in CHCl3, adsorbed onto Celite 545 (Acros Organics) and was separated with normal-phase flash chromatography (40 g silica gel column) with a 55.0-minute hexane/CHCl3/MeOH gradient at a 40 mL/min flow rate (Figure S4). This generated a total of 8 fractions. Fraction RO-5 (115 mg) went through a second stage of normal-phase flash chromatography (Figure S5), after dissolving in chloroform, absorbing onto Celite 545 (Acros Organics), with a 35.0-minute hexane/CHCl3/MeOH gradient and was separated using a 15.5 g silica gel column at a flow rate of 30 mL/min. This fractionation yielded 6 simplified fractions, in which the bioactive fraction RO-5–4 (approximately 10 mg) was purified further via preparative HPLC using a gradient system 80:20 to 90:10 of CH3CN:H2O with 0.1% formic acid over 30 minutes (Figure S6). This yielded 4 fractions: RO-5–4-1 (3.88 mg eluted at 4.0 min), RO-5–4-2 (2.27 mg eluted at 12.5 min), RO-5–4-3 (3.15 mg eluted at 15.9 min), and RO-5–4-4 (5.4 mg eluted at 0.5 min).
Khin M., Knowles S.L., Crandall W.J., Jones DD J.r., Oberlies N.H., Cech N.B, & Houriet J. (2021). Capturing the Antimicrobial Profile of Rosmarinus officinalis against Methicillin-resistant Staphylococcus aureus (MRSA) with Bioassay-guided Fractionation and Bioinformatics. Journal of pharmaceutical and biomedical analysis, 197, 113965.
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2

Cholesterol Oxidation Product Analysis

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Cholesterol, linoleic acid, oleic acid, COPs standards [7-ketocholesterol
(7-keto), 6-ketocholesterol (6-keto), 7α-hydroxy-cholesterol
(7α-OH), 7β-hydroxycholesterol (7β-OH),
5,6α-epoxycholesterol (5,6α-EP), 5,6β-epoxycholesterol
(5,6β-EP), 25-hydroxycholesterol (25-OH), 20-hydroxycholesterol (20-OH),
and cholestanetriol (triol)], butylated hydroxytoluene (BHT), pyridine, and
silicic acid (100 mesh) were purchased from Sigma-Aldrich Co., LLC (Korea).
Bis-[trimethylsilyl]-trifluoroacetamide (BSTFA)+1%
trimethylchlorosilane (TMCS) was obtained from Supelco (USA). Hexane, ethyl
acetate, acetone, methanol and chloroform of HPLC grades, celite545, and calcium
phosphate (CaHPO4·2H2O) were purchased from Fisher
Scientific Co. (USA).
Choe J., Min J.S., Lee S.O., Khan M.I., Yim D.G., Lee M, & Jo C. (2018). Influence of Cooking, Storage Period, and Re-heating on Production of Cholesterol Oxides in Chicken Meat. Korean Journal for Food Science of Animal Resources, 38(3), 433-441.
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3

Biomaterial for Stem Cell Differentiation

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Poly(ethylene glycol) (PEG; Mn 2000 Da and 3350 Da), acryloyl chloride, cystamine dihydrochloride, L-cysteine (Cys), reduced glutathione (GSH), calcium hydride, dexamethasone, β-glycerophosphate, ascorbic acid, indomethacin, isobutyl methylxanthine, insulin, pyridine, activated charcoal, and Sephadex G-25 were purchased from Sigma-Aldrich (USA). The disuccinimidyl carbonate was obtained from Acros Organics (USA). Dulbecco’s modified eagle medium (DMEM; high glucose) was purchased from Hyclone. Fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco. Anhydrous Dichloromethane (DCM), acetonitrile, anhydrous diethyl ether, dithiothreitol (DTT), tris (2-carboxyethyl) phosphine (TCEP), potassium bicarbonate (K2CO3), potassium iodide (KI), hydrochloric acid (HCl), anhydrous sodium sulphate, sodium chloride (NaCl) and Celite 545 were purchased from Fisher Scientific. DCM and acetonitrile were dried using calcium hydride. Irgacure (D2959) was procured from Ciba, Switzerland. CellTracker Red (CMTPX) and Live/Dead assay kit were obtained from Life Technologies.
Kar M., Shih Y.R., Velez D.O., Cabrales P, & Varghese S. (2015). Poly(ethylene glycol) hydrogels with cell cleavable groups for autonomous cell delivery. Biomaterials, 77, 186-197.
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4

Lipid Profiling and Oxidation Analysis

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Cholesterol, linoleic acid, oleic acid, cholesterol oxide standards [7-ketocholesterol (7-keto), 6-ketocholesterol (6-keto), 7α-hydroxycholesterol (7α-OH), 7β-hydroxycholesterol (7β-OH), 5,6α-epoxycholesterol (5,6α-EP), 5,6β-epoxycholesterol (5,6β-EP), 25-hydroxycholesterol (25-OH), 20-hydroxycholesterol (20-OH), and cholestanetriol (triol)], butylated hydroxytoluene (BHT), Xpyridine, and silicic acid (100 mesh) were purchased from Sigma-Aldrich Co., LLC (Seoul, Korea). Bis(trimethylsilyl)trifluoroacetamide (BSTFA) + 1 % trimethylchlorosilane (TMCS) was obtained from Supelco (Bellefonte, PA, USA). HPLC-grade hexane, ethyl acetate, acetone, methanol, chloroform, Celite 545, and calcium phosphate (CaHPO4.2H2O) were purchased from Fisher Scientific Co. (Malvern, PA, USA).
Khan M.I., Min J.S., Lee S.O., Yim D.G., Seol K.H., Lee M, & Jo C. (2015). Cooking, storage, and reheating effect on the formation of cholesterol oxidation products in processed meat products. Lipids in Health and Disease, 14, 89.
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5

Cholesterol Oxidation Compound Analysis

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Cholesterol, linoleic acid, oleic acid, Cholesterol oxide standards (7-keto, 6-keto, 7α-OH, 7β-OH, 5,6α-EP, 5,6β-EP, 25-OH, 20-OH, and triol), butylated hydroxytoluene (BHT), pyridine, and silicic acid (100 mesh) were purchased from Sigma-Aldrich Co., LLC (Seoul, Korea). Bis-[trimethylsilyl]-trifluoroacetamide (BSTFA) + 1 % trimethylchlorosilane (TMCS) was obtained from Supelco (Bellefonte, PA, USA). HPLC grade hexane, ethyl acetate, acetone, methanol, and chloroform, Celite 545, and calcium phosphate (CaHPO4.2H2O) were purchased from Fisher Scientific Co. (Malvern, PA, USA).
Min J.S., Lee S.O., Khan M.I., Yim D.G., Seol K.H., Lee M, & Jo C. (2015). Monitoring the formation of cholesterol oxidation products in model systems using response surface methodology. Lipids in Health and Disease, 14, 77.
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6

Cholesterol Oxidation Products Analysis

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Cholesterol, COPs standards [7-ketocholesterol (7-keto), 6-ketocholesterol (6-keto), 7α-hydroxy-cholesterol (7α-OH), 7β-hydroxycholesterol (7β-OH), 5,6α-epoxycholesterol (5,6α-EP), 5,6β-epoxycholesterol (5,6β-EP), 25-hydroxycholesterol (25-OH), 20-hydroxycholesterol (20-OH), and cholestanetriol (triol)], butylated hydroxytoluene (BHT), pyridine, and silicic acid (100 mesh) were purchased from Sigma-Aldrich Co., LLC (Korea). Bis-(trimethylsilyl)-trifluoroacetamide (BSTFA) + 1% trimethylchlorosilane (TMCS) was obtained from Supelco (Bellefonte, USA). HPLC grade hexane, ethyl acetate, acetone, methanol, and chloroform, Celite 545, and calcium phosphate (CaHPO4.2H2O) were purchased from Fisher Scientific Co. (USA).
Min J.S., Khan M.I., Lee S.O., Yim D.G., Seol K.H., Lee M, & Jo C. (2016). Impact of Cooking, Storage, and Reheating Conditions on the Formation of Cholesterol Oxidation Products in Pork Loin. Korean Journal for Food Science of Animal Resources, 36(1), 23-28.
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7

Tobacco Leaf Virus Inoculation Protocol

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Freshly infected leaves from tobacco plants were pre-frosted in liquid nitrogen and ground [tissue: buffer; 1:6 (weight/ volume)] with cold and freshly prepared potassium phosphate buffer (0.01 M), having pH of 7.0, consisting of 0.01 M 2-mercaptoethanol and 0.2% sodium sulfite, in a pre-chilled pestle and mortar. The ground inoculum was filtered through a muslin cloth to remove the debris. Celite 545 (Fisher Scientific) and Carborundum (Fisher Scientific) with a ratio of 1:2. The prepared inoculum was placed on ice till its use for inoculation of the test plants [52 (link)].
Kamran A., Li Y., Zhang W., Jiao Y., Farooq T., Wang Y., Liu D., Jiang L., Shen L., Wang F, & Yang J. (2024). Insights into the genetic variability and evolutionary dynamics of tomato spotted wilt orthotospovirus in China. BMC Genomics, 25, 40.
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8

Large-scale extraction and purification of fungal compounds

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A. fumigatus strains were inoculated (1.0×106 spores mL−1) into 1 L GMM43 (link) in a 2 L Erlenmeyer flask at 37 °C with shaking at 220 rpm. After 3.5 days, liquid fungal cultures including fungal tissue and media were frozen using a dry ice acetone bath, and lyophilized. The lyophilized residues were extracted with 500 mL of MeOH for 1 h with vigorous stirring. Extracts were filtered over cotton and evaporated on Celite-545 (Acros Organics) to dryness. The dry Celite was then loaded into a 25 gram solid phase cartridge (Isco) and subjected to large-scale reverse phase chromatography (see Chromatographic enrichment section below for details). Compounds 4 and 5 are also minor components of the culture media.
Baccile J.A., Spraker J.E., Le H.H., Brandenburger E., Gomez C., Bok J.W., Macheleidt J., Brakhage A.A., Hoffmeister D., Keller N.P, & Schroeder F.C. (2016). Plant-like biosynthesis of isoquinoline alkaloids in Aspergillus fumigatus. Nature chemical biology, 12(6), 419-424.
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9

DNA Extraction from Bacterial Cells

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Bacterial cells were inoculated into 9 ml Brain Heart Infusion (Mediaproducts BV) and grown overnight at 37°C. From this suspension, template DNA was prepared by using either whole bacterial cells without extracting DNA, or extracting DNA based on the method of Boom et al. [3] (link). Briefly, in the latter case, 0.1 ml bacterial cell suspension was added to 600 µl mixture of L6 and Celite 545 (Acros Organics, Geel, Belgium), and put at ambient temperature for 10 min. The mixture was centrifuged in an Eppendorf microcentrifuge for 10 s at 13,400×g. The pellet was washed with 70% ethanol (v/v) and subsequently acetone and centrifuged for 10 s at 13,400×g after each washing step. The acetone was disposed of after which the pellet was left to dry for 10 min at 56°C. The pellet was dissolved in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) during 10 min at 56°C.
van Tongeren S.P., Roest H.I., Degener J.E, & Harmsen H.J. (2014). Bacillus anthracis-Like Bacteria and Other B. cereus Group Members in a Microbial Community Within the International Space Station: A Challenge for Rapid and Easy Molecular Detection of Virulent B. anthracis. PLoS ONE, 9(6), e98871.
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10

Large-scale extraction and purification of fungal compounds

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A. fumigatus strains were inoculated (1.0×106 spores mL−1) into 1 L GMM43 (link) in a 2 L Erlenmeyer flask at 37 °C with shaking at 220 rpm. After 3.5 days, liquid fungal cultures including fungal tissue and media were frozen using a dry ice acetone bath, and lyophilized. The lyophilized residues were extracted with 500 mL of MeOH for 1 h with vigorous stirring. Extracts were filtered over cotton and evaporated on Celite-545 (Acros Organics) to dryness. The dry Celite was then loaded into a 25 gram solid phase cartridge (Isco) and subjected to large-scale reverse phase chromatography (see Chromatographic enrichment section below for details). Compounds 4 and 5 are also minor components of the culture media.
Baccile J.A., Spraker J.E., Le H.H., Brandenburger E., Gomez C., Bok J.W., Macheleidt J., Brakhage A.A., Hoffmeister D., Keller N.P, & Schroeder F.C. (2016). Plant-like biosynthesis of isoquinoline alkaloids in Aspergillus fumigatus. Nature chemical biology, 12(6), 419-424.
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