The polymerosome mixtures were prepared by dissolving the block PEG-PLGA (poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolide)) copolymer (molecular weight: PEG Mn 2000, PLGA Mn 4500, average Mn 6500 (total); lactide:glycolide ratio: 65:35; Sigma-Aldrich, Darmstadt, Germany) at a 10 mg/mL concentration in a chloroform solution (Sigma-Aldrich, Darmstadt, Germany). The chloroform was removed completely by evaporation. A thin and fine polymer film on the glass was obtained. The film was further dried under a constant flow of Nitrogen 3.5 (Messer Poland Ltd., Chorzow, Poland) for 3 min. Subsequently, the film was rehydrated with solutions of Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, Darmstadt, Germany) containing 10% DMSO (purity ≥ 99.9%; Sigma-Aldrich, Darmstadt, Germany). The vehicles were fabricated with a mix of the solution. To homogenize the sample, the suspension was subjected to sonication (UP50H, Hielscher, Teltow, Germany). The blank sample was prepared the same way, only it did not contain DMSO.
Chloroform solution
Manufactured by Merck Group
Sourced in United States
Chloroform solution is a clear, colorless liquid that is commonly used as a solvent in various laboratory applications. It has a dense, heavy nature and a characteristic sweet odor. This product is suitable for numerous chemical and analytical procedures where a stable, high-purity solvent is required.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Trioctylamine, by Merck Group (2 mentions)
Oleic acid, by Merck Group (2 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Up50h, by Hielscher (1 mentions)
Hematoxylin solution, by Abcam (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Eosin y solution, by Merck Group (1 mentions)
Follicle dpc growth medium, by PromoCell (1 mentions)
Dulbecco s phosphate buffer saline dpbs, by Corning (1 mentions)
Rotary evaporator, by Heidolph (1 mentions)
Up50h ultrasonic processor, by Hielscher (1 mentions)
Milli q, by Merck Group (1 mentions)
Iron acetylacetonate, by Merck Group (1 mentions)
5 protocols using chloroform solution
1
Polymerosome Fabrication and Characterization
2
Evaluation of Androgenic Activity in Hair Follicles
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Dimethyl sulfoxide (DMSO), 100× penicillin-streptomycin, chloroform solution, and eosin Y solution were purchased from Sigma-Aldrich (Saint Louis, MO, USA). A hematoxylin solution and a dihydrotestosterone enzyme-linked immunosorbent assay (ELISA) kit were purchased from Abcam, Inc. (Cambridge, MA, USA). Dulbecco’s phosphate-buffer saline (DPBS) was purchased from Corning, Inc. (Corning, NY, USA). DMEM, 0.25% Trypsin-EDTA, and TRIzol were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Follicle DPC growth medium was purchased from PromoCell GmbH (Heidelberg, Germany). EZ-Cytox cell viability assay reagent was purchased from DoGenBio, Inc. (Seoul, Republic of Korea). HelixCript Thermo Reverse Transcriptase and RNase inhibitor were purchased from Nanohelix, Inc. (Daejeon, Republic of Korea). A TB Green® Premix Ex Taq kit was purchased from Takara Korea Biomedical, Inc. (Seoul, Republic of Korea). Collagen type I-coated wells were purchased from SPL, Inc. (Gyenggi, Republic of Korea). Pansidil was purchased from Dongkook Pharmaceutical, Inc. (Seoul, Republic of Korea).
3
Niosomal Amikacin Encapsulation by Thin-Film Hydration
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The niosomes containing amikacin were synthesized based on the thin-film hydration method (Thabet et al. 2022 (link)). In this method, the weight amounts of span 60 and tween 60 with cholesterol are dissolved in chloroform solution (all from Sigma Aldrich, USA) and evaporated under vacuum at 60 ℃ by rotary evaporator (1 h, 120 rpm) (Heidolph Instruments, Germany). Next, hydration of the formed film was performed using 1 mg/ml of amikacin (Tehran-Darou, Iran) solution in phosphate buffer (PBS) (pH 7.2, one h, 120 rpm). Finally, the compositions prepared for 7 min were sonicated using a probe sonicator (Hielscher up50H ultrasonic processor, Germany). The samples were kept at 4 ℃ for further investigations.
4
Synthesis of PSMA-Coated Iron Oxide Nanoparticles
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Fe3-δO4 iron oxide nanoparticles (IONPs) were synthesized using thermal decomposition, as previously described (Huang et al., 2011 (link)). Briefly, 1.42 g of iron acetylacetonate was mixed with 0.57 mL of oleic acid and 20 mL of trioctylamine (all 3: Sigma-Aldrich, St. Louis, MO, United States). The solution was refluxed at 325°C in an argon environment for 30 min. The solution was then cooled to room temperature and washed with a toluene-ethanol solution. A magnet was used to concentrate and collect the NPs, which were then transferred to chloroform solutions (Merck, Whitehouse Station, NJ, United States) and mixed with 0.4 mg/mL of prostate specific membrane antigen (PSMA) (Sigma-Aldrich) for 6 h at 55°C. Finally, the PSMA-coated NPs were collected and washed 3 times with high purity water (MQ water, 18.2 MΩ cm, Milli-Q; Merck Ltd., Taipei, Taiwan).
5
Synthesis of Fe3-δO4 Nanoparticles
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Fe3-δO4 nanoparticles (22 nm and 14 nm) were synthesized using thermal decomposition, as previously described30 (link). Briefly, 1.42 g of iron acetylacetonate was mixed with 0.57 mL of oleic acid and 20 mL of trioctylamine (all 3: Sigma-Aldrich, St. Louis, MO). The solution was refluxed at 325 °C in an argon environment for 30 minutes. After the solution had cooled down to room temperature, the precipitates were collected with a magnet and washed 3 times with toluene. The Fe3-δO4 nanoparticles were collected with a magnet and then transferred to chloroform solutions (Merck, Whitehouse Station, NJ) containing 0.4 mg/mL of PSMA (Sigma-Aldrich) and were left there for 6 hours at 55 °C. The Fe3-δO4 nanoparticles were collected, washed 3 times with MQ water, and then stored at 4 °C.
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