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Pa5 92370

Manufactured by Thermo Fisher Scientific
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Sourced in United States, United Kingdom

The PA5-92370 is a laboratory equipment product. It serves a core function in scientific research and analysis, but a detailed description while maintaining an unbiased and factual approach is not available.

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Lab products found in correlation

Sta 6000, by PerkinElmer (1 mentions) Ht7700, by Hitachi (1 mentions) Phosphotungstic acid, by Merck Group (1 mentions) Zetaview pmx 110, by Particle Metrix (1 mentions) Ab125011, by Abcam (1 mentions) Ab18529, by Abcam (1 mentions) Af5139, by Affinity Biosciences (1 mentions) C digit blot scanner, by LI COR (1 mentions) Amersham ecl prime western blotting detection reagent, by Cytiva (1 mentions) Ib401002, by Thermo Fisher Scientific (1 mentions) Na9340, by Merck Group (1 mentions) Tbs t, by Merck Group (1 mentions) Ab22595, by Abcam (1 mentions) Ab175396, by Abcam (1 mentions) Gm130, by Thermo Fisher Scientific (1 mentions) Amersham ecl prime, by Cytiva (1 mentions)

5 protocols using pa5 92370

1

Antibody-Functionalized Magnetic Nanoparticles

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GMNPE was generated with GMNPs (Fe3O4@SiO2-PEG-CHO) bound by the rabbit antibody to CD63 (PA5-92,370, Thermo Fisher Scientific). In short, 1 mg/mL GMNP solution was purified by magnetic separation, and incubated with anti-CD63 antibody overnight at 4 °C to obtain GMNPE. Fluorescent GMNPE was prepared using the same method, with the aforementioned GMNPs replaced by RhB-labeled GMNPs.
In order to assess drug carrying capacity of GMNPE, thermogravimetric analysis (TGA; STA 6000, PerkinElmer) was conducted for analyzing MNPs, GMNPs, and GMNPE. The adsorption capacity and stability of HSA were measured with FITC-labeled HSA by fluorescence spectroscopy.
Xu C., Wang Z., Liu Y., Wei B., Liu X., Duan K., Zhou P., Xie Z., Wu M, & Guan J. (2022). Extracellular vesicles derived from bone marrow mesenchymal stem cells loaded on magnetic nanoparticles delay the progression of diabetic osteoporosis via delivery of miR-150-5p. Cell Biology and Toxicology, 39(4), 1257-1274.
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2

Comprehensive Exosome Characterization Protocol

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For morphological analysis, exosomes were fixed with 2.5% glutaraldehyde and placed on carbon-coated copper grids for 5 min. The sample was then stained with 2% phosphotungstic acid (Sigma‒Aldrich, USA) for 3 min. Images were captured by transmission electron microscopy (TEM; HT7700, Hitachi, Japan). Exosome number and size were assessed by nanoparticle tracking analysis (NTA) using a ZetaView PMX 110 (Particle Metrix, Germany). The surface markers of the exosomes were verified by Western blot analysis. The following antibodies were used: anti-CD9 (1:1000, AF5139, Affinity Biosciences), anti-CD63 (1:1000, PA5-92370, Thermo Fisher Scientific), anti-TSG101 (1:1000, ab125011, Abcam) and anti-Lamp2b (1:500, ab18529, Abcam).
Lin Z., Xu G., Lu X., Liu S., Zou F., Ma X., Jiang J., Wang H, & Song J. (2024). Chondrocyte-targeted exosome-mediated delivery of Nrf2 alleviates cartilaginous endplate degeneration by modulating mitochondrial fission. Journal of Nanobiotechnology, 22, 281.
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3

Western Blot Analysis of Extracellular Vesicles

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For Western blot, proteins separated by SDS-PAGE starting from EV lysates were transferred onto a PVDF membrane using semi-dry transfer at 8 mA for 1.5 h. Transfer buffer consisted of 48 mM Tris base, 39 mM glycine, 1 mM SDS and 20% (v/v) methanol. Membranes were either directly tested or stored at −20 °C in 5% milk in PBS. Alix was detected with the commercial monoclonal antibody 3A9 (diluted 1:500, Thermo Fisher Scientific, Waltham, MA, USA) and CD63 with rabbit polyclonal antibodies (diluted 1:1000, PA5-92370, Thermo Fisher Scientific) both in combination with HRP-conjugated secondary antibodies.
Neumair J., D’Ercole C., De March M., Elsner M., Seidel M, & de Marco A. (2023). Macroporous Epoxy-Based Monoliths Functionalized with Anti-CD63 Nanobodies for Effective Isolation of Extracellular Vesicles in Urine. International Journal of Molecular Sciences, 24(7), 6131.
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4

Immunofluorescence Analysis of CD63

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GMNPBSA and GMNPE were fixed in 4% paraformaldehyde, blocked, and probed with rabbit antibody to CD63 (PA5-92,370, Thermo Fisher Scientific). The cells were re-probed with goat anti-rabbit (#60,839, 1:50, Cell Signaling Technologies, Beverly, MA) conjugated to Alexa Fluor® 555. Finally, images were captured under an 80i fluorescence microscope.
Xu C., Wang Z., Liu Y., Wei B., Liu X., Duan K., Zhou P., Xie Z., Wu M, & Guan J. (2022). Extracellular vesicles derived from bone marrow mesenchymal stem cells loaded on magnetic nanoparticles delay the progression of diabetic osteoporosis via delivery of miR-150-5p. Cell Biology and Toxicology, 39(4), 1257-1274.
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5

Extracellular Vesicle Protein Analysis

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Small EV (5 µg total protein) were separated by sodiumdodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE) on a 4–20% gradient gel (4561094, Bio-Rad, Hercules, CA, USA), transferred to PVDF membranes (IB401002, Thermo Fisher Scientific Inc.), and blocked with 5% non-fat dry milk dissolved in Tris-buffered saline containing 0.05% Tween 20 (Merck KGaA; TBST). The following proteins were analysed: CD63 (1:500, PA5-92370, Thermo Fisher Scientific Inc.), CD81 (1:1000, PA5-79003, Thermo Fisher Scientific Inc.), CD73 (1:1000, ab175396, Abcam, Cambridge, United Kingdom), Calnexin (1:1000, ab22595, Abcam) and GM130 (1:1000, MA5-35107, Thermo Fisher Scientific Inc.). Horseradish peroxidase-coupled donkey anti-rabbit antibody (NA9340, Merck KGaA) was used as secondary antibody in a 1:1000 dilution. The binding of the antibodies was detected using the chemiluminescent Amersham ECL Prime Western Blotting Detection Reagent (GEHERPN2232, Cytiva, Marlborough, MA, USA) on a C-DiGit Blot Scanner (LI-COR Biosciences, Lincoln, NE, USA).
Tscherrig V., Cottagnoud S., Haesler V., Renz P., Surbek D., Schoeberlein A, & Joerger-Messerli M.S. (2023). MicroRNA Cargo in Wharton’s Jelly MSC Small Extracellular Vesicles: Key Functionality to In Vitro Prevention and Treatment of Premature White Matter Injury. Stem Cell Reviews and Reports, 19(7), 2447-2464.
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