Sephadex g25 column pd 10

Manufactured by Cytiva

Sephadex G25 column (PD-10) is a pre-packed size exclusion chromatography column used for desalting, buffer exchange, and sample cleanup. It is filled with Sephadex G-25 resin, which separates molecules based on their size and molecular weight. The column has a cut-off molecular weight of approximately 5,000 Daltons, allowing the separation of small molecules from larger biomolecules such as proteins, peptides, or nucleic acids.

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Lab products found in correlation

Dioleoylphosphatidylserine dops, by Avanti Polar Lipids (1 mentions)

2 protocols using sephadex g25 column pd 10

Preparation of SapC-DOPS Nanovesicles

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CellVue Maroon (CVM; Molecular Targeting Technologies Inc., Exton, PA) in ethanol was mixed with DOPS and SapC for bath sonication preparation as described previously [9 (link)]. Briefly, 82 μg DOPS (Avanti Polar Lipids, Alabaster, AL) were mixed with 30 μl CVM (from 1 mM stock) in a glass tube and the mix solvent evaporated under nitrogen gas. Human recombinant SapC protein [47 (link)] (0.4 mg) was added to DOPS-CVM along with 20 μl citrate/phosphate buffer pH 5.0. After addition of 1 ml PBS (or DMEM) and a 30 min sonication step, SapC-DOPS nanovesicles were separated from free CVM dye using a Sephadex G25 column (PD-10; Amersham Pharmacia Biotech, Piscataway, NJ). DOPS-CVM nanovesicles were prepared as described by omitting SapC.

Blanco V.M., Chu Z., Vallabhapurapu S.D., Sulaiman M.K., Kendler A., Rixe O., Warnick R.E., Franco R.S, & Qi X. (2014). Phosphatidylserine-selective targeting and anticancer effects of SapC-DOPS nanovesicles on brain tumors. Oncotarget, 5(16), 7105-7118.

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Preparation of Fluorescent SapC-DOPS Nanovesicles

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Dioleoylphosphatidylserine (DOPS; Avanti Polar Lipids Inc., Alabaster, AL) was dried under a stream of N₂ gas, combined with recombinant SapC protein (27 (link)) in citrate/phosphate buffer (pH 5.0) and bath sonicated in PBS as described in detail previously (21 (link)). To fluorescently label SapC-DOPS nanovesicles, an aliquot of CellVue® Maroon (CVM, Molecular Targeting Technology Inc., Exton, PA) in ethanol was dried together with DOPS before SapC addition and bath sonication. Free CVM was eliminated from this preparation by filtration through a Sephadex^™ G25 column (PD-10, Amersham Pharmacia Biotech, Piscataway, NJ). For in vivo treatment, a lyophilized formulation was used: dry DOPS was suspended in 80% tert-butanol. SapC and sucrose (10 mg/ml) were dissolved in water. DOPS suspension and SapC plus sucrose (1:0.6, vol:vol) was lyophilized in a freeze dryer (VirTis Unitop 1000L linked to a Freezemobile 25XL; The VIRTIS, Gardiner, NY). The stable powder cake was resuspended in saline solution to form SapC-DOPS nanovesicles. Vesicles were monitored with a submicron particle size analyzer (Coulter Model N4 Plus; Fullerton, CA). The molar ratio of SapC:DOPS was 1:7.

Zhao S., Chu Z., Blanco V.M., Nie Y., Hou Y, & Qi X. (2015). SapC-DOPS Nanovesicles as Targeted Therapy for Lung Cancer. Molecular cancer therapeutics, 14(2), 491-498.

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