Conjugation of streptavidin (Santa Cruz, Dallas, TX, USA) with gold nanoparticles was prepared according to the methods described previously with minor modifications [27 (link),28 (link)]. Prior to use, the pH value of gold nanoparticle solution was adjusted to 7.0 by using 0.2 M sodium carbonate and 0.1 M hydrochloric acid. Two hundred microliters of streptavidin was added to 20 mL of gold nanoparticle solution. The mixture was stirred with a magnetic stirrer for 30 min at room temperature in the dark (Schott bottle was wrapped with aluminum foil), followed by the addition of 200 µL 10% bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA) and 800 µL of 2% polyethylene glycol 3000 (Sigma-Aldrich, St. Louis, MO, USA). The mixture was incubated for 1 h at room temperature. After that, the mixture was centrifuged for 30 min at 20,000× g, 4 °C. The supernatant containing unbound streptavidin was removed as much as possible. The pellet (consisting of conjugated streptavidin) was resuspended with 500 µL of 0.02% polyethylene glycol 3000, 0.05% sodium azide (Sigma-Aldrich, St. Louis, MO, USA). The gold-conjugated streptavidin was then stored at 4 °C.
Polyethylene glycol 3000
Manufactured by Merck Group
Sourced in United States, Germany
Polyethylene glycol 3000 is a water-soluble, non-toxic, and inert polymer. It has a molecular weight of approximately 3,000 Daltons. Polyethylene glycol 3000 is commonly used in various laboratory applications as a solvent, emulsifier, or stabilizer.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Sodium azide, by Merck Group (2 mentions)
Peg 3000, by Merck Group (2 mentions)
Polyethylene glycol 4000, by Merck Group (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Ethanol absolute for analysis, by Merck Group (1 mentions)
Chitosan, by Merck Group (1 mentions)
Tween 80, by Thermo Fisher Scientific (1 mentions)
Ninhydrin, by Merck Group (1 mentions)
Span 80, by Merck Group (1 mentions)
Hydrindantin, by Merck Group (1 mentions)
High molecular weight chitosan, by Merck Group (1 mentions)
Lithium acetate anhydrous, by Thermo Fisher Scientific (1 mentions)
Tween 80, by Merck Group (1 mentions)
Pexidartinib, by Selleck Chemicals (1 mentions)
Pexidartinib, by Merck Group (1 mentions)
Safranin o, by Merck Group (1 mentions)
Gellan gum, by Merck Group (1 mentions)
6 protocols using polyethylene glycol 3000
1
Conjugation of Streptavidin with Gold Nanoparticles
2
Conjugation of Streptavidin with Gold Nanoparticles
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Conjugation of streptavidin (Santa Cruz, Dallas, TX, USA) with gold nanoparticles was prepared according to the methods described previously with minor modifications [27 (link),28 (link)]. Prior to use, the pH value of gold nanoparticle solution was adjusted to 7.0 by using 0.2 M sodium carbonate and 0.1 M hydrochloric acid. Two hundred microliters of streptavidin was added to 20 mL of gold nanoparticle solution. The mixture was stirred with a magnetic stirrer for 30 min at room temperature in the dark (Schott bottle was wrapped with aluminum foil), followed by the addition of 200 µL 10% bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA) and 800 µL of 2% polyethylene glycol 3000 (Sigma-Aldrich, St. Louis, MO, USA). The mixture was incubated for 1 h at room temperature. After that, the mixture was centrifuged for 30 min at 20,000× g, 4 °C. The supernatant containing unbound streptavidin was removed as much as possible. The pellet (consisting of conjugated streptavidin) was resuspended with 500 µL of 0.02% polyethylene glycol 3000, 0.05% sodium azide (Sigma-Aldrich, St. Louis, MO, USA). The gold-conjugated streptavidin was then stored at 4 °C.
3
Celecoxib Formulation Development
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Celecoxib (CCX, Mw = 381.37 g/mol) and magnesium stearate (lubricant, Mw = 591.27 g/mol) were purchased from Fagron Nordic A/S (Copenhagen, Denmark). Polyethylene glycol 3000 (PEG 3000, Mw ≈ 3000 g/mol) and polyethylene glycol 4000 (PEG 4000, Mw ≈ 4000 g/mol) were kindly gifted by Merck (Darmstadt, Germany). All chemicals were used as received.
4
Chitosan-Based Nanoparticle Formulation
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Low-molecular-weight chitosan (molecular weight: 20 000–50 000 Da, degree of deacetylation ≥ 90%; Zhejiang Aoxing Biotechnology Co. Ltd., China) and high-molecular-weight chitosan (molecular weight: 310 000–375 000 Da, degree of deacetylation ≥ 75.0%; Sigma-Aldrich, Ireland) were used as the matrix material of nanoparticles with glacial acetic acid as the solvent of chitosan (Merck, Germany). Tween 80 (Fisher Scientific, UK), span 80 (Merck, Germany) and ethanol absolute for analysis (Merck, Germany) were used as additives. Lithium acetate anhydrous (ACROS Organics™, USA), ninhydrin and hydrindantin (Sigma-Aldrich, USA) were used in quantification assay of chitosan. Lactose monohydrate (Sorbolac 400; Meggle, Germany) was used as the microparticulate carrier with polyethylene glycol 3000 (PEG3000; Merck, Germany) as the stabilizer.
5
Pexidartinib Depletion of Macrophages
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The CSF1R inhibitor pexidartinib (Selleck Chemicals) was used to deplete macrophages. pexidartinib was initially suspended and stored at 200 mg/mL in DMSO, and further suspended in 5% DMSO, 45% polyethylene glycol 3000 (Sigma-Aldrich), 5% Tween-80 (Sigma-Aldrich), and 45% ddH2O for a working concentration of 10 mg/mL. FVB/N mice bred in-house at 5 weeks of age were given a working solution of pexidartinib at 45 mg/kg once daily by oral gavage for 2 weeks. Mammary glands were then either fixed in 4% paraformaldehyde for histology, processed for flow cytometry or lysed for HA analysis by ELISA as previously described (Bohrer et al., 2014 (link)). HA concentrations were normalized to mammary gland weight.
6
Bacterial Cellulose Hydrogel Synthesis
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Bacterial cellulose (BC) was prepared at our laboratory at Tomas Bata University in Zlin. Gellan gum (GG, low acyl gellan), polyethylene glycol 3000 (PEG), crystal violet (C25N3H30Cl, purity ≥90%), safranin O (C20H19N3N4Cl, purity ≥85%) and sodium hydroxide (NaOH) were supplied by Sigma-Aldrich (Darmstadt, Germany). Citric acid (CA, C6H8O7) as green crosslinker and hydrochloric acid (HCl) were purchased from Penta s.r.o. (Zlin, Czech Republic). The reagents were of analytical grade with high purity and were used without further purification. Distilled water was used for the preparation of all aqueous solutions. Waste whey was collected from Kromilk A.S (Kromeriz, Czech Republic). Black tea was purchased from a grocery store in Zlin, Czech Republic. Sucrose was supplied by Amersco LLC (MA, USA). All reagents were used without further purification.
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