The synthesis of 18F-fallypride (Fig-1 ) was carried out in the chemical process control unit (CPCU) using modifications of previously reported methods (Mukherjee et al., 1995, 2002 (link)). Purification of 18F-fallypride was carried out by high performance liquid chromatography (HPLC) separation on an Econosil reverse-phase C-18 semi-prep column 250 × 10 mm; 10μm particle size (Alltech Assoc., Deerfield, IL). Eluents used were 60% acetonitrile and triethylamine (0.1%) in water at a flow rate of 2.5 ml/min in a Gilson Gradient System consisting of one UV detector with wavelength fixed at 280 nm and a radiation flow detector. The radioactive fraction appearing at 20 min was collected, solvents removed and the residue was taken up in sterile saline and passed through a Whatman 0.22-μm filter and dispensed for in vitro and in vivo studies in specific activity was 74 GBq/μmol with a radiochemical purity of >98%.
0.22 μm filter
Manufactured by Cytiva
Sourced in United Kingdom
The 0.22 μm filter is a sterile, single-use filter designed for the removal of microorganisms from liquids. It features a 0.22 μm pore size membrane that effectively retains bacteria and other particulates while allowing the passage of the desired liquid.
Automatically generated - may contain errors
Lab products found in correlation
55fecl3, by PerkinElmer (2 mentions)
Nanosight lm10, by Malvern Panalytical (1 mentions)
Ix73 light microscope, by Olympus (1 mentions)
Uv vis detector, by Thermo Fisher Scientific (1 mentions)
Dionex asi 100, by Thermo Fisher Scientific (1 mentions)
P680 hplc pump, by Thermo Fisher Scientific (1 mentions)
Pda 100 photodiode array detector, by Thermo Fisher Scientific (1 mentions)
15 protocols using 0.22 μm filter
1
Synthesis and Purification of 18F-Fallypride
2
Cultivation and Stress Response of Nori Algae
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Artificial sterilized seawater (SEALIFE, Marinetech, Tokyo, Japan) enriched with ESS2 [71 (link)] was used for the maintenance and examination of the three life stages of N. yezoensis (strain U-51)—gametophytes, sporophytes, and conchosporophytes [51 (link)]—under 60–70 μmol m–2 s–1 of light with a short-day photoperiod (10 h light/14 h dark) at 15 °C. Air was filtered through a 0.22 μm filter (Whatman, Maidstone, UK). The culture medium was changed weekly.
Heat stress and pharmacological treatments were performed as described by Khoa and Mikami [50 (link)]. In brief, the three life stages were incubated at 5 °C, 15 °C, or 25 °C for 0.5, 1, 2, 4, 6, 8, and 12 h. The experiments started at 12:00, 3 h after the start of light irradiation (9:00). In the short-day photoperiod with 10 h light, the 8 and 12 h time points comprised 1 and 5 h of darkness, respectively. Continuous 24 h lighting was also employed as a control condition. Treatments with 2.5 mM of benzyl alcohol (BA) and 4% dimethyl sulfoxide (DMSO) were performed by incubating algae at 15 °C for 5, 15, or 30 min. Algal cells were also treated with BA at 25 °C in the light or with DMSO under dark conditions for 8 or 12 h. After these treatments, algae were frozen in liquid nitrogen and stored at −80 °C prior to their use for gene expression analysis.
Heat stress and pharmacological treatments were performed as described by Khoa and Mikami [50 (link)]. In brief, the three life stages were incubated at 5 °C, 15 °C, or 25 °C for 0.5, 1, 2, 4, 6, 8, and 12 h. The experiments started at 12:00, 3 h after the start of light irradiation (9:00). In the short-day photoperiod with 10 h light, the 8 and 12 h time points comprised 1 and 5 h of darkness, respectively. Continuous 24 h lighting was also employed as a control condition. Treatments with 2.5 mM of benzyl alcohol (BA) and 4% dimethyl sulfoxide (DMSO) were performed by incubating algae at 15 °C for 5, 15, or 30 min. Algal cells were also treated with BA at 25 °C in the light or with DMSO under dark conditions for 8 or 12 h. After these treatments, algae were frozen in liquid nitrogen and stored at −80 °C prior to their use for gene expression analysis.
3
Radiolabeling of Desferrated cMBT
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55Fe-cMBT was prepared according to Ryndak et al.5 (link). 200 μM Fe-cMBT in H2O was mixed with 50 mM EDTA pH 4 in a 1:1 volume ratio. The solution was incubated overnight at room temperature while shaking. The solution was filtered with a 0.22 μM filter (Whatman) and desferrated cMBT was extracted with one volume of chloroform. The chloroform phase was washed twice with one volume of H2O and evaporated. Desferrated cMBT was taken up in 100 % EtOH and 0.4 mCi 55FeCl3 (Perkin-Elmer) was added. Remaining desferrated cMBT was saturated by dropwise addition of cold FeCl3 until no further increase in red color was observed. 55Fe-cMBT was purified again by extraction with one volume of chloroform and washed twice with one volume of H2O to remove free 55Fe3+ and Fe3+.
4
Isolation and Characterization of ADSC-derived Exosomes
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The exosomes derived from ADSCs were purified from serum-free cell medium by series centrifugation and filtration steps. Briefly, collected supernatant centrifuged at 300 g for 10 min to remove dead cells, 5,000 g for 10 min to remove cellular debris. Then the supernatant was ultrafiltrated by 0.22 μm filter (Whatman, Maidstone, United Kingdom) and centrifuged at 120,000 g for 90 min. The final pellets were resuspended in 1 mL PBS and stored at −80°C.The collected pellets were distinguished by transmission electron microscopy (TEM, Zeiss, Axio, Germany) and nanoparticle tracking analysis (NTA, NanoSight LM10, Malvern Instruments, Westborough, MA).
5
Stability of Trastuzumab-Doxorubicin Conjugate
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Example 32
Human plasma (prepared using heparin) was centrifuged at 200 g for 10 minutes at 4° C. and the supernatant was incubated with protein A agarose (Kem-En-Tec) for 1 hour at 4° C. to deplete for IgG. The depleted plasma was filter sterilized using a 0.22 μm filter (Whatman). The trastuzumab-doxorubicin conjugate derived from 28a was added to the sterile human plasma to a final concentration of 100 μg/ml and incubated at 37° C. in a CO2 incubator to keep plasma pH levels close to the physiological pH of 7.2. Samples were taken at 0, 24, 48 and 144 hours and stored at −80° C. The trastuzumab conjugate was purified with protein A agarose followed by MS analysis. Human plasma samples were incubated with protein A agarose for 1 hour at 4° C., washed three times with phosphate-buffered saline (PBS) and the trastuzumab conjugate was eluted with 100 mM glycine-HCl pH 2.7 followed by neutralization with 1 M Tris-HCl pH 8.0. MS analysis showed that the peak corresponding to the trastuzumab-doxorubicin conjugate (50708 Da) did not decrease in time. In addition, no peaks corresponding to degradation products could be detected, proving that the conjugate is stable for at least 144 hours in human plasma.
6
Culturing Filamentous Marine and Freshwater Bangia
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Gametophytes of filamentous Bangia grown on rocks were harvested on April 20, 2018 from Kamomejima Island (41°52′ N, 140°06′ E) in Esashi, Hokkaido in Japan. The thalli were maintained in sterilized artificial seawater (SEALIFE, Marinetech, Tokyo, Japan) enriched with ESS2 [29 (link)] under 60–70 μmol photons m−2 s−1 light with a short-day photoperiod (10 h light/14 h dark) at 15 °C with air filtered through a 0.22 μm filter (Whatman, Maidstone, UK). Conchocelis filaments appeared during the culture of thalli and were maintained as described for thalli; the conchosporangia parasitically developed on the conchocelis filaments. The culture medium was changed weekly. Thalli of the freshwater species Bangia atropurpurea, which were collected from the rocky bed of a river with a rapid current in Higashi-kawachisawa, Shizuoka, Japan in May 2005 and April 2006, were maintained according to Yokono et al. [17 (link)] in commercially available Ca2+-rich mineral water (Contrex®, Nestlé Waters Marketing & Distribution) in plastic culture vessels, except that the culture conditions described above for marine Bangia species were utilized. Thalli, conchocelis, and conchosporangia were observed and imaged under an Olympus IX73 light microscope (Olympus, Tokyo, Japan) equipped with an Olympus DP22 camera.
7
Radiolabeling of Desferrated cMBT
8
Simulating Acid and Alkali Spill Impact on Soil
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Considering the frequency of chemical accidents and frequency of use, HCl (35%, Daejung, Korea) and NaOH (98%, Daejung, Korea) were selected as strong acid and alkali, respectively [1 (link),22 ]. We simulated an extreme but plausible acid or alkali spill situation that acid or alkali chemical was continuously spilled from a storage tank at a plant for a few days. For the experiment, 10 g of the untreated soil were placed in a 50-mL conical tube, and 30 mL of 10 M HCl or NaOH were added. The reaction was conducted in a rotating shaker at 25 °C and 40 rpm for two days, then the suspension was centrifuged, and the supernatant solutions were filtered through a 0.22-μm filter (Whatman, UK). The separated soils were washed with deionized water five times to remove excess salts and dissolved ions. Because excess H+ and OH− remaining after washing could affect the titration experiment, HNO3 (60%, Daejung, Korea) or NaOH was added to the washed soils until the supernatant pH reached a range of pH 6–8. The suspensions were centrifuged and decanted, and the residual soil was washed five times with deionized water and freeze-dried. The physicochemical properties of the acid- and alkali-spilled soils are summarized in Table S2 .
9
Platelet-Free Plasma Isolation
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Blood samples were taken in ethylenediaminetetraacetic acid tubes (BD, Plymouth, UK) and within 1 h centrifuged for 10 min at 1690 g by 4°C to obtain plasma. Plasma was centrifuged at 10,000 g for 30 min by 4°C to obtain platelet free plasma (pfp). Thereafter the supernatant was passed through a 0.22 μm filter (Whatmann) and aliquoted. Aliquots were stored at −80°C until EV isolation.
10
Cultivation of Bangia sp. ESS1 Gametophyte
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Filamentous gametophytic thalli of ‘Bangia’ sp. ESS1 were harvested in Esashi, Hokkaido, Japan on May 17, 2010 (Hirata et al., 2011 (link)) and phylogenetically classified as a member of ‘Bangia’ group 2 (Li et al., 2019a (link)). The alga was maintained clonally as an experimental line in our laboratory in sterilized artificial seawater as described by Li et al. (2019b) (link) under 60–70 μmol photons m−2 s−1 light with a short-day photoperiod (10 h light/14 h dark) at 15°C and aerated with air filtered through a 0.22-μm filter (Whatman, Maidstone, UK). The culture medium was changed weekly.
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