Sephadex g 50 medium

Manufactured by Merck Group

Sourced in United States

Sephadex G-50 medium is a gel filtration medium designed for the separation and purification of biomolecules. It is a cross-linked dextran-based material that functions as a molecular sieve, allowing the separation of molecules based on their size and shape. The medium is commonly used in applications such as desalting, buffer exchange, and the purification of proteins, peptides, and nucleic acids.

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Sephadex G-50 (101 citations)

7 protocols using sephadex g 50 medium

Quantifying Insulin Degradation via Elution

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¹²⁵I-TyrA14-insulin degradation was determined by elution profile from a Sephadex G-50 (medium, Sigma-Aldrich) column as previously established [34 (link)]. Briefly, samples were loaded onto a 50 cm column and eluted with 0.15 M ammonium bicarbonate buffer, 0.02% BSA and 5% sodium azide, pH 8.0. Column elution fractions were quantified on a gamma counter, adjusted for efficiency, and expressed as disintegrations per min (DPM). To verify that the column elution profile discriminates intact from degraded insulin, ¹²⁵I-TyrA14-insulin (200 pmol/l) was incubated with or without HepG2 cells overnight at 37°C. ¹²⁵I-TyrA14-insulin samples collected after transcytosis assays were compared with native and HepG2-degraded insulin elution peaks.

Gray S.M., Aylor K.W, & Barrett E.J. (2017). Unravelling the regulation of insulin transport across the brain endothelial cell. Diabetologia, 60(8), 1512-1521.

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Liposomal Dox Formulation Preparation

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DOX hydrochloride was provided by Huafeng Lianbo Technology Co. Ltd. (Beijing, People’s Republic of China). Hydrogenated soy phosphatidylcholine (HSPC; MW 785) was obtained from Lucas Meyer GmbH (Düsseldorf, Germany). Cholesterol (CH) was supplied by Nanjing Xinbai Pharmaceutical Co. Ltd. (Nanjing, People’s Republic of China). 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-(methoxy[polyethylene glycol]-2000) (mPEG₂₀₀₀-DSPE) was purchased from J&K Scientific (Beijing, People’s Republic of China). Sephadex^® G-50 (medium) was obtained from Sigma-Aldrich (St Louis, MO, USA). All other reagents were of analytical grade.

Song Y., Huang Z., Song Y., Tian Q., Liu X., She Z., Jiao J., Lu E, & Deng Y. (2014). The application of EDTA in drug delivery systems: doxorubicin liposomes loaded via NH4EDTA gradient. International Journal of Nanomedicine, 9, 3611-3621.

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Liposomal Doxorubicin Formulation Development

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Citric acid (99.8%) and ethylenediamine (≥99%) were purchased from Sigma–Aldrich Ltd. (Poole, UK). The phospholipids 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-glycerol sodium salt (DPPG) were obtained from Lipoid GmbH (Ludwigshafen, Germany), while 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] ammonium salt (DSPE-PEG) was obtained from Avanti Polar Lipids (Alabaster, AL, USA). Doxorubicin hydrochloride (DOX) was kindly donated by Regulon SA (Athens, Greece). Nucleopore filters of 100 nm pore size (Whatman, Maidstone, UK) were employed for liposome extrusion. Sephadex G-50 (medium) and thiazolyl blue tetrazolium bromide (MTT) were obtained from Sigma–Aldrich (St. Louis, MA, USA). Cell culture RPMI 1640 medium with L-glutamine, fetal bovine serum (FBS), penicillin/streptomycin, phosphate buffer saline (PBS), and trypsin (0.05% w/v) /EDTA (0.25% w/v) were purchased from Invitrogen Ltd. (Paisley, UK). All other reagents and solvents were of analytical grade and used without further purification.

Mavroidi B., Kaminari A., Sakellis E., Sideratou Z, & Tsiourvas D. (2023). Carbon Dots–Biomembrane Interactions and Their Implications for Cellular Drug Delivery. Pharmaceuticals, 16(6), 833.

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Lipid and Peptide Synthesis Protocol

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1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPG) and dodecylphosphocholine (DPC) were purchased from Avanti Polar Lipids (USA). Rink amide polystyrene resin and amino acid derivatives for peptide synthesis were from Iris Biotech GmbH (Germany); trifluoroacetic acid (TFA, analytical and HPLC grades), triisopropylsilane and 2,2,2-trifluoroethanol from Sigma-Aldrich (USA); N,N’-diisopropylcarbodiimide from Fluka (Germany); 1-hydroxybenzotriazole and 1,2-etanoditiol from Nova Biochem-Merck (Germany); N,N-dimethylformamide, diisopropyl ether, chloroform and dichloromethane were obtained from Vetec (Brazil); and acetonitrile (HPLC grade) from JT Baker (USA). Sodium dodecyl sulfate (SDS), calcein, Sephadex® G-50 medium, Triton X-100, and HEPES from Sigma-Aldrich (USA). Unless stated otherwise analytical grade solvents were used.

Gusmão K.A., dos Santos D.M., Santos V.M., Cortés M.E., Reis P.V., Santos V.L., Piló-Veloso D., Verly R.M., de Lima M.E, & Resende J.M. (2017). Ocellatin peptides from the skin secretion of the South American frog Leptodactylus labyrinthicus (Leptodactylidae): characterization, antimicrobial activities and membrane interactions. The Journal of Venomous Animals and Toxins Including Tropical Diseases, 23, 4.

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Antioxidant Characterization of Lipid Vesicles

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All chemicals were purchased at the highest purity available and were used without further purification. Ethanol, Sephadex G-50 medium, cholesterol, curcumin, oleic acid, the reagent grade salts for the 50 mM K–phosphate and 100 mM KCl (pH 7.0) buffer solutions, 2,20-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), and sodium cholate (SC) were purchased from Sigma-Aldrich. Lipoid S100 (LS100) was from Lipoid (Lipoid GmbH, Ludwigshafen, Germany). According to the manufacturer’s information, LS100 is constituted of approximately 100% soybean phosphatidylcholine. Eudragit S100 was a kind gift of Evonik (Evonik Industries AG, Darmstadt, Germany). Sterile filters of cellulose acetate (0.2 µm) were from Advantec. All aqueous solutions were prepared by using water obtained from a Milli-Q gradient A-10 system (Millipore, Burlington, MA, USA, 18.2 MΩ cm, organic carbon content ≤ 4 μg/L).

De Leo V., Milano F., Mancini E., Comparelli R., Giotta L., Nacci A., Longobardi F., Garbetta A., Agostiano A, & Catucci L. (2018). Encapsulation of Curcumin-Loaded Liposomes for Colonic Drug Delivery in a pH-Responsive Polymer Cluster Using a pH-Driven and Organic Solvent-Free Process. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(4), 739.

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Enzymatic Colorimetric Assays for Biomolecules

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Hydrogen peroxide 30 wt.% (VWR Chemicals), horseradish peroxidase ~150 U/mg (77332, Sigma-Aldrich), absolute ethanol (VWR Chemicals, ≥99.8), microcrystalline cellulose
(BASF Basionics, ≥95%), DMSO (Alfa Aesar, ≥99%), glucose oxidase from Aspergillus niger (G7141, Sigma-Aldrich) (AnGOx), bovine serum albumin (05470, Sigma-Aldrich, ≥96%), Sodium iodide (383112, Sigma-Aldrich, ≥99.5%), sodium phosphate dibasic (S9763, Sigma-Aldrich, ≥99%), sodium phosphate monobasic (S3139, Sigma-Aldrich), sodium carbonate (S7795, Sigma-Aldrich, ≥98%), sodium bicarbonate (S5761, Sigma-Aldrich, ≥99.5%), sodium sulfite (S0505, Sigma-Aldrich, ≥98%), Phenol (328111, Sigma-Aldrich, ≥99%), Amplex ® Red reagent (Invitrogen), 2,6-DichloroindoPhenol (D1878, Sigma-Aldrich), Sephadex G-50 Medium (Sigma-Aldrich), 3,5-Dinitrosalicylic acid (D0550, Sigma-Aldrich, ≥98%), Coomassie Brilliant Blue G-250 staining (Sigma, Deisenhofen, Germany), fluorescein isothiocyanate (F7250, Sigma-Aldrich, ≥90%).

Califano D., Kadowaki M.A.S., Calabrese V., Prade R.A., Mattia D., Edler K.J., Polikarpov I, & Scott J.L. (2020). Multienzyme Cellulose Films as Sustainable and Self-Degradable Hydrogen Peroxide-Producing Material. Biomacromolecules, 21(12).

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Lipid-based Nanoformulation Preparation

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All chemicals were purchased of the highest purity available and were used without further purification. Ethanol, methanol, Sephadex G-50 medium, cholesterol, beclomethasone dipropionate (BDP), the reagent grade salts for the 50 mM K-phosphate, 100 mM KCl (pH 7.0) buffer solutions and sodium cholate were purchased from Sigma-Aldrich. 1,2-Dipalmitoyl-sn-glycero-3-
ammonium salt (LissRhod PE) were purchased from Avanti Polar Lipids. Lipoid S100 (LS100) was from Lipoid (Lipoid GmbH, Germany). According to the manufacturer's instructions, LS100 is constituted by approximately 100% soybean phosphatidylcholine (PC). Pluronic F127 ®
[poly(ethylene oxide) -poly(propylene oxide) -poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer, PF127] was a kind gift of BASF (Ludwigshafen, Germany). Sterile filters of cellulose acetate of 0.2 µm were from Advantec. All aqueous solutions were prepared by using water obtained from a Milli-Q gradient A-10 system (Millipore, 18.2 MΩ cm, organic carbon content ≤4 μg/L).

De Leo V., Ruscigno S., Trapani A., Di Gioia S., Milano F., Mandracchia D., Comparelli R., Castellani S., Agostiano A., Trapani G., Catucci L, & Conese M. (2018). Preparation of drug-loaded small unilamellar liposomes and evaluation of their potential for the treatment of chronic respiratory diseases. International journal of pharmaceutics, 545(1-2).

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