Wha230600

Manufactured by Cytiva

The WHA230600 is a laboratory equipment product. It is designed for use in scientific and research environments. The core function of this product is to perform specific tasks or procedures related to laboratory work. No further details about its intended use or capabilities are provided.

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Lab products found in correlation

Wha110604, by Cytiva (2 mentions) Nanodrop 2000c, by Thermo Fisher Scientific (2 mentions) Biotinyl cap pe, by Avanti Polar Lipids (1 mentions)

2 protocols using wha230600

Synthesis of Biotinylated Liposome SUVs

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SUVs were prepared using a 10 mL LIPEX Extruder (Transferra Nanosciences, Inc.). Lipids were mixed in ~500 μL chloroform with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) (850375C, Avanti Polar Lipids) as the base lipid. Biotinylated lipids were incorporated at 0.05–0.2 mol% 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(biotinyl) (Biotinyl-Cap PE) (870282C, Avanti Polar Lipids). To directly tag the membrane in control experiments, N-(fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (FITC DHPE) (23304, AAT Bioquest) was incorporated at 0.1 mol%. Lipids were dried first by rotary evaporation and second by ultrahigh purity N₂. Lipid cakes were resuspended and sonicated in 3 mL nanopure water (final concentration, 2 mg mL⁻¹) prior to performing three freeze-thaw cycles. SUVs in nanopure water were then extruded 10× through a 0.08 μm polycarbonate filter (WHA110604, Whatman) supported by a drain disc (WHA230600, Whatman). SUVs were used within ~2 weeks.

Glazier R., Brockman J.M., Bartle E., Mattheyses A.L., Destaing O, & Salaita K. (2019). DNA mechanotechnology reveals that integrin receptors apply pN forces in podosomes on fluid substrates. Nature Communications, 10, 4507.

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Preparation of Fluorescent SUV Liposomes

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Small unilamellar vesicles (SUV) were prepared by extrusion. 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) (850375C, Avanti Polar Lipids) and Texas Red 1,2Dihexadecanoyl-sn-Glycero-3-Phosphoethanolamine, Triethylammonium Salt (TR DHPE) were mixed in ~100 μL chloroform at 100 mol % DOPC or 99.5 mol% DOPC and 0.5 mol % TR DHPE. Chloroform was evaporated by rotary evaporation, and lipid cakes were dried under vacuum and under ultra-high purity nitrogen. Lipids were resuspended in water at 2 mg/mL and sonicated for 30 s prior to 3 freeze-thaw cycles. SUVs were extruded 10x in a 10 mL LIPEX Extruder (Transferra Nanosciences, Inc.) fit with a 0.08 μm polycarbonate filter (WHA110604, Whatman) and a drain disc (WHA230600, Whatman). The final concentration of TR in liposomes was measured on a Thermo Scientific Nanodrop 2000c spectrophotometer.

Brockman J.M., Su H., Blanchard A.T., Duan Y., Meyer T., Quach M.E., Glazier R., Bazrafshan A., Bender R.L., Kellner A.V., Ogasawara H., Ma R., Schueder F., Petrich B.G., Jungmann R., Li R., Mattheyses A.L., Ke Y, & Salaita K. (2020). Live-cell super-resolved PAINT imaging of pN cellular traction forces. Nature methods, 17(10), 1018-1024.

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