Oil in water (O/W) emulsion (oil to water 20:75% w/w) and whey protein isolate (WPI) concentration of 0.5% (w/w) was prepared by dissolving WPI in saline buffer (150 mM NaCl and 0.02% w/w NaN3). The mixture was stirred with a magnetic stirrer until dissolution. Sunflower oil, which was previously treated with florisil (Taufkirchen, Sigma, F9127), was used as the oil phase [75 (link)]. The mixture of sunflower oil and protein dispersion was further vortexed for 3 min to obtain a coarse emulsion. The pre-emulsion was sonicated with an ultrasound generator (Sonics VCX 500, Sonics & Materials Inc., Newtown, CT, USA) with a 0.13 cm diameter titanium probe with an amplitude of 80%, pulse duration of 5 s on/10 s off for 3 min. Lipolysis results were carried out on split samples, one-half for each bile salt.
Vcx 500
Manufactured by Sonics
Sourced in United States
The VCX 500 is a versatile laboratory equipment designed for a variety of applications. It features a compact and durable construction, making it suitable for use in various research and testing environments. The core function of the VCX 500 is to provide controlled and consistent sample processing capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Kta fplc system, by GE Healthcare (3 mentions)
Cve 3100, by Eyela (2 mentions)
Mpeg nh2, by Abbexa (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Qubit 2.0 fluorometer, by Thermo Fisher Scientific (2 mentions)
Florisil, by Merck Group (1 mentions)
Jsm 7500f, by JEOL (1 mentions)
Icps 7500, by Shimadzu (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Nisin, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Desk 4, by Denton (1 mentions)
Mannose agarose resin, by Merck Group (1 mentions)
Vivaspin 20, by Sartorius (1 mentions)
Histrap hp column, by GE Healthcare (1 mentions)
Freezone 2.5 freeze dryer, by Labconco (1 mentions)
N 1 filter paper, by Cytiva (1 mentions)
Ethanol, by Merck Group (1 mentions)
Malvern zetasizer nano zs90, by Malvern Panalytical (1 mentions)
5 l bioreactor, by Infors (1 mentions)
65 protocols using vcx 500
1
Preparation and Sonication of O/W Emulsion
2
Characterization of Nanomaterial Synthesis
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The following equipment was used: ultrasonicactor (SONICS-VCX500, SONICS, Newtown, CT, USA), centrifuge (UNIVERSAL 320R, Hettich, Beverly, MA, USA), field emission scanning electron microscope (FESEM, JSM-7500F, JEOL, Tokyo, Japan), vacuum muffle furnace (Neytech Qex, DEGUSSA-NEY DENTAL, INC., Bloomfield, CT, USA), light source (FOK-100W, Fiber Optic Korea, Cheonan, Korea), photodetectivity measurement system (4200-SCS, Keithley, Beaverton, OR, USA), thermal evaporation system, and solar simulator for the on/off test (XES-301S, SAN-EI ELECTRIC CO., Ltd., Osaka, Japan).
3
Magnetotactic Bacteria-Derived Magnetic Nanoparticles
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MNPs (Fe3O4), obtained from an anaerobically cultured magnetotactic bacterium (Magnetospirillum sp. AMB-1, American Type Culture Collection), were used to generate 3D hESCs, called hEBs. The magnetic bacterium was cultured in a modified magnetic spirillum growth medium, as previously described [48 (link),50 (link)]. In brief, bacteria were cultivated via a fermenter (FMT-ST-S05, Fermentec) in an anaerobic condition for 5 d at 27 °C. Then, the bacterial cells were centrifuged at 11,300 × g for 20 min, and then the collected cells were sonicated with 35% amplification for 15 min (VCX500, Sonics & Materials). The MNPs were isolated from bacterial cell debris using 100-mm petri dishes with neodymium-iron-boron (NdFeB) magnets attached below. Only MNPs, except for cell debris, were attached along the perimeter of the magnets. Therefore, the MNPs were collected while the cell debris was washed out. The separated MNPs were washed 3 times using phosphate-buffered saline (PBS; Welgene), relying on magnetic adhesion. Then, the MNPs were sterilized using an autoclave. After measuring the concentration of iron ions using inductively coupled plasma atomic emission spectroscopy (ICPS-7500, Shimadzu), the MNPs were stored in a concentration of 1 mg/ml in PBS at 4 °C. Before use, the MNPs were dispersed using bath type ultrasonicator (JAC 1002, Kodo Technical Research) for 10 min.
4
Conductive PEDOT:PSS-PGCL Composite Films
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PEDOT:PSS aqueous solution (PH1000, Clevious, USA) was subjected to liquid nitrogen (N2) for 10 min and then placed under vacuum conditions (5 mTorr) to lyophilize at − 85 °C for 24 h. The resulting solid (1 wt%) was dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich, USA) solvent for 2 h using an ultrasonic processor (VCX-500, Sonics and Materials, USA). Different ratios of 1 wt% PGCL (55:45) solution (in DMSO solvent) and d -sorbitol were mixed with the PEDOT:PSS solution to evaluate the changes in mechanical and electrical performance of the conductive composite. The composite solutions were drop-cast onto a PDMS mold and dried in an oven at 60 °C for 2 days to create conductive PGCL films. The conductivity of the composite was measured using a 4-point probe system (Keithley 2400 digital multimeter, Keithley). The stretchability of the composite was evaluated using a universal testing machine (Instron 8801, Intron).
5
Nisin-Induced Expression of PFK and GK in L. lactis
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To research the expression levels of PFK and GK in Lactococcus lactis NZ9000, nisin (Sigma-Aldrich, St. Louis, MO) was used to express the protein; the specific steps are as follows. We cultured Lactococcus lactis NZ9000-pNZ8148-PFK, Lactococcus lactis NZ9000-pNZ8148-GK, and Lactococcus lactis NZ9000-pNZ8148 in GM17 medium supplemented with 10 μg/mL chloramphenicol at 30°C. Cultures were stimulated with 10 ng/mL nisin at an optical density at 600 nm of 0.5 and cultured for 12 h. We centrifuged for 5 min (12,000 × g) to harvest the cells at 4°C. Cell pellets were washed twice in ice-cold 1× PBS (pH 7.2) and resuspended in PBS (Thermo Fisher). Ultrasound was used to break the cells for 30 cycles of 5 s at -2°C (VCX 500, Sonics and Materials Inc., Newtown, CT). We centrifuged (12,000 × g) for 5 min at 4°C to remove cell debris, to obtain cell extracts. Bovine serum albumin (Thermo Fisher) was used as the standard protein, and the protein concentration was determined according to the Bradford method. For protein analysis of samples, the cell extracts were mixed with 5-fold concentrated SDS-PAGE loading buffer after heating at 100°C for 5 min. Finally, we performed SDS-PAGE on a 12% (wt/vol gel, with 20 uL of sample per well.
6
Nanoemulsion Formulation and Characterization
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Phase diagrams were constructed by using the water titration method to recognize the type of structure for emulsification and to typify the behavior of the mixtures with dilution paths. The blank oil-in-water (O/W) NE was prepared under optimal conditions using the water titration method followed by probe sonication. NE was prepared by adding 4% (v/v) of Frankincense oil, 36% (v/v) of pre-warmed Smix i.e. Tween 20: Transcutol P (3 : 1), and 60% double distilled water. The ratio between oils to surfactants (O/S) was maintained at 1: 9, as this formulation exhibited optimal stability. The resulting mixture was then vortexed for approximately 5 min at room temperature followed by heating and sonication until a clear and transparent solution is obtained (Sonics VCX500 (U.S.A)). The sonication was carried out for approximately 20 min until a clear and transparent phase formed indicating the production of blank nanoemulsion (B-NE). Each experiment was performed in triplicate (n = 3). Furthermore, B-NE was incorporated with a combination of active components (ER + PTX) in 1:1 to produce EPNE.
7
Fluorometric Assay for Protease Activity
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Example 7
Tumor and muscle tissue samples from PyMT 8119 tumor bearing mice are collected and frozen at −80° C. The tissues are thawed and homogenized in cold TCNB buffer (pH 7.5, 50 mM Tris-HCl, 10 mM CaCl2, 150 mM NaCl and 0.05% Brij35) at 100 mg/200 μL using ultrasonic disruption (VCX500, Sonics & Materials Inc, Newtown, Conn.). After homogenates are centrifuged at 15,000 g at 4° C. for 20 min, supernatants are collected. APMA (p-aminophenylmercuric acetate, 90 μL, 2 mM in TCNB buffer) is added to the supernatants (90 μL). The resulting mixtures are incubated at 37° C. for 1 h before use. 500 nM of SDM-42 is used for the cleavage of 45 μL of activated tissue supernatants (final volume: 50 μL). The assay is carried out using a SpectraMax M2 spectrometer with SoftMax Pro v4.5 software. Fluorescence signals of (λex, 620 nm, λem, 670 nm), (λex, 620 nm, λem, 773 nm) and (λex, 720 nm; λem, 773 nm), where λex and λem stand for excitation and emission wavelengths respectively, are measured as a function of time at room temperature. Samples are measured in triplicate.
8
Preparation and Characterization of SWCNT-Coated Filter Paper
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Example 2
The SWCNTs suspension was prepared in accordance with Cui et al. (See Hu, L. B. et al., Proc. Natl. acad. Sci. USA 200, 106, 214900-21494 and Hu, L. B., et al., Nano Lett 2010, 10, 708-714.) Briefly, 3.0 mg/mL of SWCNTs were dispersed in a 1.0 mg/mL of SDBS solution and sonicated for 3 hours using a tip sonicator (Sonics and Materials VCX 500 VibraCell, Newton, Conn.) in an ice bath to avoid overheating. A 6.5×5.5 cm qualitative filter paper was coated with the SWCNTs suspension using a conventional paintbrush. It was then dried at room temperature for 10 minutes and then in an oven at 60° C. for 20 minutes. The conductivity of the resulting coated filter paper was measured using a source-measurement unit (Keithley Source Meter Model 2400, Cleveland, Ohio). This process of coating, drying, and measuring the conductivity of the filter paper was considered one cycle. The filter paper was treated for six cycles. Images of the topography of the filter paper before and after coating with SWCNTs were obtained using Zeiss Ultra 55 Scanning Electron Microscopy (SEM). To obtain SEM images of the filter paper without coating, gold sputtering (Denton Vacuum LLC Desk IV, Moorestown, N.J.) was performed on it to give necessary conductivity to the filter paper.
9
Efficient Purification of Recombinant PHL Protein
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E. coli Tuner (DE3)/pET25b_phl cells were grown in standard LB broth low-salt medium (ForMedium, UK) containing 100 μM ampicillin at 37°C until the OD600 reached ~ 0.5. After induction with 0.2 mM isopropyl ß-d -1-thiogalactopyranoside (ForMedium, UK), cells were cultured for an additional 20 hours at 18°C, harvested by centrifugation at 12,000 g for 10 min and resuspended in buffer A (300 mM NaCl, 20 mM Tris/HCl, pH 7.5). Harvested cells were stored at -20°C prior to protein purification.
Cells were disrupted by sonication (VCX 500, Sonics & Materials, Inc., USA) and the soluble fraction was collected by centrifugation at 21,000 g at 4°C for 1 hour and filtrated through a 0.45 μm pore size filter (Carl Roth, Germany). Recombinant protein PHL was purified by affinity chromatography on mannose-agarose resin (Sigma-Aldrich, USA) equilibrated with buffer A using an ÄKTA FPLC system (GE Healthcare, UK). The protein was eluted isocratically. Protein purity was assessed by SDS-PAGE (12% gel) stained with Coomassie Brilliant Blue R-250/G-250 (Sigma-Aldrich, USA). The fractions containing pure PHL were extensively dialyzed against an appropriate buffer and used for further studies. If desired, PHL was concentrated using an ultrafiltration unit with a 10-kDa cut-off membrane (Vivaspin 20, Sartorius, Germany).
Cells were disrupted by sonication (VCX 500, Sonics & Materials, Inc., USA) and the soluble fraction was collected by centrifugation at 21,000 g at 4°C for 1 hour and filtrated through a 0.45 μm pore size filter (Carl Roth, Germany). Recombinant protein PHL was purified by affinity chromatography on mannose-agarose resin (Sigma-Aldrich, USA) equilibrated with buffer A using an ÄKTA FPLC system (GE Healthcare, UK). The protein was eluted isocratically. Protein purity was assessed by SDS-PAGE (12% gel) stained with Coomassie Brilliant Blue R-250/G-250 (Sigma-Aldrich, USA). The fractions containing pure PHL were extensively dialyzed against an appropriate buffer and used for further studies. If desired, PHL was concentrated using an ultrafiltration unit with a 10-kDa cut-off membrane (Vivaspin 20, Sartorius, Germany).
10
Recombinant TRP Protein Expression
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The open reading frame (ORF) of gene trp (LmjF.31.2010) was amplified by PCR using the following primers containing the restriction enzyme sites for BamHI and NotI (underlined): TRP_F_BamHI 5´-GGATCC ATGGAGTGCATCAACCAAGAGAGC-3´ and TRP__R_NotI 5´-GCGGCCGC TCACATGGCACAGATAAACACC-3´. The amplified fragment was cloned into the pET28a (Novagen, USA) expression vector and sequenced to confirm the insertion direction. The pET-TRP plasmid obtained was then used to transform into E. coli (BL21(DE3)CodonPlus-RIL). Selected clones were grown aerobically at 37°C in LB medium containing kanamycin (30 μg/mL) and chloramphenicol (35 μg/mL) to a culture OD600 0.6–0.8. pET-TRP expression was induced by 1 mM of isopropyl-β-D-thiogalactopyranoside. After induction, the culture was lysed by sonication (Sonics–VCX500) with 20 mM sodium phosphate, 500 mM sodium chloride and 5 mM imidazole. Lysed samples were clarified by centrifugation at 10,000 x g, for 15 minutes at 4°C, and inclusion bodies were solubilized with 20 mM sodium phosphate, 500 mM sodium chloride, 5 mM imidazole and 8 M urea. Recombinant TRP was obtained by affinity chromatography with a 1 mL HisTrap HP column (GE Healthcare, Uppsala, Sweden). The purified recombinant TRP was analyzed by SDS-PAGE and then used to produce a rabbit polyclonal anti-TRP antibody by Proteimax Biotechnology (Sao Paulo, Brazil).
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