Zeolite granules CaA and NaX (granule size 1.6–2.5 mm, Luoyang Jalong Micro-nano New Materials Co., Ltd., Henan, China), calcium chloride (anhydrous, 93% purity, Alfa Aesar) and strontium chloride (anhydrous, 99% purity, Alfa Aesar) were purchased and used as pristine materials. To reduce the formation of unexpected salts, e.g., NaCl, during the impregnation process, CaA and NaX granules were first treated by ion exchange to replace the Ca2+ and Na+ cations with Sr2+. The impact of time on the ion-exchange process and the concentration of the SrCl2 solution during the ion-exchange process were investigated (Section S1, ESI† ). All the ion-exchanged granules were dried at 150 °C for 1 h. As-received CaA granules were directly impregnated with CaCl2 solution since the formation of byproduct salts was not expected. The impregnation process was done by dripping the AEMHs solution to the granules as shown in Fig. 1(f) . The loading of the AEMHs solution was from 12 wt% to 45 wt% (Section S2, ESI† ). After the impregnation, the granules were dried at 150 °C for 1 h. The processing conditions of the obtained impregnated granules are listed in Table 1 .
Calcium chloride
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, India, China, Thailand, Germany, Canada, Australia, Belgium
Calcium chloride is a chemical compound with the formula CaCl2. It is a white crystalline solid that is highly soluble in water. Calcium chloride is commonly used as a desiccant, a road de-icing agent, and in various industrial and laboratory applications.
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Lab products found in correlation
Sodium chloride (nacl), by Thermo Fisher Scientific (22 mentions)
Magnesium chloride, by Thermo Fisher Scientific (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Potassium chloride (kcl), by Thermo Fisher Scientific (9 mentions)
Trypsin edta, by Thermo Fisher Scientific (9 mentions)
Sodium bicarbonate, by Thermo Fisher Scientific (9 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (7 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Chitosan, by Merck Group (6 mentions)
Sodium alginate, by Merck Group (6 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (5 mentions)
Urea, by Thermo Fisher Scientific (5 mentions)
Antibiotic antimycotic solution, by Thermo Fisher Scientific (4 mentions)
Gentamycin, by Thermo Fisher Scientific (4 mentions)
Citric acid, by Thermo Fisher Scientific (4 mentions)
Acetonitrile, by Thermo Fisher Scientific (4 mentions)
Calcium nitrate tetrahydrate, by Thermo Fisher Scientific (3 mentions)
Sodium hydrogen carbonate, by Thermo Fisher Scientific (3 mentions)
Magnesium chloride hexahydrate, by Thermo Fisher Scientific (3 mentions)
Sodium sulfate, by Thermo Fisher Scientific (3 mentions)
88 protocols using calcium chloride
1
Cation-exchanged Zeolite Granules for AEMHs
2
Synthesis and Characterization of Biochemical Compounds
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Sodium hypophosphite hydrate (NaH2PO2·H2O, 98%), phosphorous acid (H3PO3, 98%), sodium phosphite dibasic pentahydrate (Na2HPO3·5H2O), adenosine (C10H13N5O4, 98%), and deuterium oxide (D2O, 99.8% atom % D) were from Acros Organic; Uridine (C9H12N2O6, 98%), standard compounds, e.g., Uridine-5-monophosphate (5′-UMP) and adenosine-5-monophosphate (5′-AMP) were from Sigma Aldrich, urea, thiourea, kaolinite clay, calcium sulphate dihydrate (CaSO4·2H2O), magnesium chloride, sodium chloride, ammonium carbonate, ammonium chloride from TCI, calcium chloride (CaCl2, 98%), white sand (SiO2) and ferrous chloride tetrahydrate (FeCl2·4H2O, 98%), and instant ocean were from Alfa Aesar. Deionized water was obtained in-house using a Barnstead (Dubuque, IA, USA) NANO pure® Diamond Analytical combined reverse osmosis-deionization system [24 (link),25 (link),26 (link)].
3
aPTT Clotting Time Inhibition Assay
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Serially diluted GAG samples (25 μL) were incubated with pooled, normal human citrated plasma (50 μL; Technoclone, Surrey, UK) and Pathromtin SL reagent (50 μL; Siemens, Erlangen, Germany) for 2 mins at 37 °C prior to the addition of calcium chloride (25 μL, 50 mM; Alfa Aesar, Heysham, UK). The time taken for clot formations to occur (an upper maximal of 2 mins was imposed, represented as 100% inhibition of clotting) was recorded using a Thrombotrak Solo coagulometer (Axis-Shield). HPLC-grade H2O (0% inhibition of clotting, representing a normal aPTT clotting time, of ≈ 37–40 seconds) and porcine mucosal heparin (193 IU mg−1; Celsus, Cincinnati, OH, USA) were screened as controls. The EC50 values of all test and control samples were determined using a sigmoidal dose response curve fitted with Prism 7 (GraphPad Software, San Diego, CA, USA).
4
Synthesis and Characterization of Polymer Materials
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Ethylenediamine (EDA), triethyl phosphate (TEP), and methyl methacrylate (MMA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). PEEK 450G was purchased from Victrex. Potassium phosphate dibasic trihydrate, sodium chloride, sodium sulfate, and sodium bicarbonate were all purchased from Arcos Organics. Tris (hydroxymethyl) aminomethane and calcium chloride were obtained from Alfa Aesar (Haverhill, MA, USA). A 1 step PNPP substrate solution was purchased from Thermofisher Scientific (catalog number 37621). The Invitrogen CyQUANT cell proliferation assay kit was purchased from Thermofisher Scientific (catalog number V13154) (Waltham, MA, USA). All materials were stored according to the manufacturer’s instructions.
5
Ultrasonic Synthesis of Nanomaterials
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Calcium chloride (CaCl 2 , 93%, Alfa Aesar), sodium carbonate monohydrate (Na 2 CO 3 • H 2 O, ≥ 99.5%, Sigma-Aldrich), sodium hexametaphosphate ((NaPO 3 ) 6 , Alfa Aesar), tetraethyl orthosilicate (TEOS, 98.0%, Aldrich), ethanol (EtOH, ≥ 99.8%, Sigma-Aldrich), ammonium hydroxide (NH 4 OH, 30.0%, Carlo Erba), hexadecyltrimethylammonium bromide (CTAB, >98.0%, Tokyo Chemical Industry Co., Ltd.), hydrochloric acid (HCl, 36% w/w, Alfa Aesar), sodium hydroxide (NaOH, 97%, Thermo Scientific), potassium hydrogen phosphate (K 2 HPO 4 , 98%, Alfa Aesar) and potassium dihydrogen phosphate (KH 2 PO 4 , 98%, Alfa Aesar) were used without additional purification.
An ultrasonic cup horn (19.5 kHz, Danacamerini prototype) was used to prepare nano-CaCO 3 and HMSNs, as shown in Fig. 1. The ultrasonic generator was used to regulate ultrasonic power, which can be adjusted up to 300 W (electric input power). The ultrasonic transducers consist of high-efficiency pre-stressed piezoelectric (PZT) rings (planar PZT Morgan Electronics, diameter 50 mm) compressed between two erga discs. The maximum capacity of the cup horn is 80 mL, which is cooled with cooling water [38] . The ultrasonic powers delivered were determined calorimetrically with water as heating media and all ultrasonic power mentioned below refers to the power measured calorimetrically [39] .
An ultrasonic cup horn (19.5 kHz, Danacamerini prototype) was used to prepare nano-CaCO 3 and HMSNs, as shown in Fig. 1. The ultrasonic generator was used to regulate ultrasonic power, which can be adjusted up to 300 W (electric input power). The ultrasonic transducers consist of high-efficiency pre-stressed piezoelectric (PZT) rings (planar PZT Morgan Electronics, diameter 50 mm) compressed between two erga discs. The maximum capacity of the cup horn is 80 mL, which is cooled with cooling water [38] . The ultrasonic powers delivered were determined calorimetrically with water as heating media and all ultrasonic power mentioned below refers to the power measured calorimetrically [39] .
6
In Vitro Microsomal Metabolism Assay
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BEX was obtained from LC Laboratories (Woburn, MA, USA). Linoleic acid was purchased from Acros Organics (Loughborough, UK). Trizma maleate, MgCl2, KH2PO4, K2HPO4, and reduced nicotinamide adenine dinucleotide phosphate (NADPH), porcine pancreatin powder (8 × USP specifications), sodium taurocholate (NaTc), NaCl, lecithin, tetrabromo-o-cresol and sunflower oil were from Sigma (Gillingham, UK). Calcium chloride was from Alfa Aesar (Lancashire, UK). Pooled male rat liver microsome was purchased from Gibco (Paisley, UK). Polyethylene glycol 400 (PEG400) was obtained from Fisher Scientific (Loughborough, UK). All solvents used were of HPLC grade or higher.
7
Macrophage Uptake of PEGylated ANS
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Macrophages were prepared in a 50 ml conical tube with 1 × 106 cell/10 ml of culture medium concentration. After overnight incubation, macrophages were detaching to the plastic, all of the media exchanging to 10 ml of fresh culture media with 50 μl of PEGylated ANS colloid (2.8 × 1011 particles/ml). ANS-MAs were rinsed three times by Hanks' Balanced Salt Solution (HBSS) with calcium chloride and magnesium chloride (Gibco, Carlsbad, CA) to remove excess noningested ANS. ANS-MAs were detached by using TrypLE Express (Invitrogen, Carlsbad, CA). ANS-MAs were confirmed by phase contrast microscopy and holographic images and the ANS location identified in the macrophage by the gold ion detection with scanning electron microscopy (SEM, JSM-7610F, JEOL, Akishima, Japan).
8
Immortalized Human Foreskin Keratinocyte Culture
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Immortalized human foreskin keratinocytes (HFK), provided by Michael Underbrink (University of Texas Medical Branch, Galveston, TX, USA), were grown in EpiLife medium (Gibco, Gaithersburg, MD, USA), supplemented with 60 µM calcium chloride (Gibco), human keratinocyte growth supplement (Gibco), and 1% penicillin-streptomycin (Caisson, Smithfield, UT, USA). U2OS and HCT116 cells were maintained in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin. Zeocin (Alfa Aesar, Ward Hill, MA, USA) and H2O2 were used to induce DSBs. NU7441 (Selleckchem) was used to inhibit DNA-PKcs phosphorylation. KU55933 (Selleckchem, Houston, TX, USA) was used to inhibit ATM kinase activity.
9
Isolation of Mononuclear Cells from Primate Samples
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PBMCs were isolated from whole blood anticoagulated with heparin (132 Units per 8 ml blood) (BD Biosciences, Oxford, UK) using standard methods. Of note is that the material used for density gradient centrifugation was adjusted dependent on the macaque species, with a Ficoll Histopaque gradient (GE Healthcare, USA) used with rhesus macaque blood and a Percoll gradient (GE Healthcare) used with cynomolgus macaques. Mononuclear cells (MNC) were isolated from spleen and lung tissue samples using an OctoMACS tissue dissociation device (Miltenyi Biotec). Lung tissue samples were dissected into approximately 5mm3 pieces and incubated for 1 h in a solution of 772.8 U/ml collagenase + 426 U/ml DNase (both from Sigma) diluted in Earle’s balanced salt solution supplemented with 200 mg/ml Calcium Chloride (Gibco, Life Technologies, Renfrew, UK), at 37 °C with continual gentle mixing of the tube. The homogenised solution was passed through a 70 µm cell filter (BD Biosciences) and the mononuclear cells separated by Ficoll Histopaque density gradient centrifugation. PBMCs and MNC isolated from tissues were stored at −180 °C until resuscitated for analysis.
10
Colonic Supernatant Generation and Utilization
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Colonic supernatants were generated as previously described.37 (link),88 (link) Briefly, full circumference colonic tissue sections of 5 mm from all mice used for somatic and visceral pain experiments were transferred and incubated for 1 hour in 96-well plate containing 300 μL Hanks’ balanced salt solution (HBSS) supplemented with calcium chloride and magnesium chloride (Gibco, Carlsbad, CA) in a humidified incubator at 37°C under 95% air and 5% CO2; samples were not standardized to the weight of segments. Supernatants were filtered (0.2 μm pore polyethersulfone membrane) and stored at −80°C. For in vitro experiments involving calcium imaging, pooled supernatants derived from mice in the same treatment group were used. Pooled supernatants from 2 animals of the appropriate treatment group were used for calcium imaging for each experiment, where naive DRGs were imaged from 2 mice at a time. Three total experiments were performed; naive DRGs from a total of 6 mice were used for the calcium imaging. These DRGs were incubated with pooled supernatants derived from a total of 6 different animals from the appropriate treatment group.
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