Sodium hyaluronate (Biohyalo 12, weight-average molecular weight 1100–1600 kDa) was obtained from SHISEIDO Co., Ltd. (Tokyo, Japan). MgCl2 and KCl were purchased from FUJIFILM Wako Pure Chemical Corporation (Tokyo, Japan). CaCl2, AlCl3, Dulbecco’s phosphate buffered saline (D-PBS) (−) were purchased from NACALAI TESQUE, INC. (Kyoto, Japan). NaCl was purchased from JUNSEI CHEMICAL Co., Ltd. (Tokyo, Japan). 5-fluoresceinamine was purchased from Sigma (St. Louis, MO, USA). 4-(4, 6-dimethoxy-1, 3, 5-triazin-2-yl)-4-methylmorpholinlum chloride (DMT-MM) was purchased from Kokusan Chemical Co., Ltd. (Tokyo, Japan). Deionized water was used as the solvent in all experiments unless otherwise stated.
Cacl2
Manufactured by Nacalai Tesque
Sourced in Japan
Calcium chloride (CaCl2) is a common laboratory chemical used as a drying agent, flocculant, and electrolyte. It is a white, crystalline solid that is highly soluble in water. CaCl2 has a core function of absorbing water and increasing the ionic strength of solutions.
Automatically generated - may contain errors
Lab products found in correlation
Potassium chloride (kcl), by Nacalai Tesque (9 mentions)
Nahco3, by Nacalai Tesque (9 mentions)
K2hpo4 3h2o, by Nacalai Tesque (7 mentions)
Na2so4, by Nacalai Tesque (7 mentions)
Mgcl2 6h2o, by Nacalai Tesque (5 mentions)
Sodium chloride (nacl), by Nacalai Tesque (4 mentions)
Tris hydroxymethyl aminomethane ch2oh 3cnh2 and 1 m hcl, by Nacalai Tesque (3 mentions)
Potassium chloride (kcl), by Fujifilm (2 mentions)
Tris hydroxymethylaminomethane ch2oh 3cnh2, by Nacalai Tesque (2 mentions)
1 m hcl, by Nacalai Tesque (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Collagenase 4, by Thermo Fisher Scientific (2 mentions)
Bone morphogenetic protein 4 (bmp4), by R&D Systems (2 mentions)
Mgcl 6h2o, by Nacalai Tesque (2 mentions)
Bovine serum albumin solution, by Merck Group (2 mentions)
Kh2po4, by Nacalai Tesque (2 mentions)
Mgcl2, by Nacalai Tesque (2 mentions)
Sodium chloride (nacl), by Fujifilm (2 mentions)
Sodium chloride (nacl), by Junsei (1 mentions)
Dulbecco s phosphate buffered saline d pbs, by Nacalai Tesque (1 mentions)
19 protocols using cacl2
1
Synthesis of Fluorescent Hyaluronic Acid Conjugates
2
Cardiomyocyte Isolation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eight weeks after the 5/6Nx operation, the heart was exposed and perfused with 10 mL of PBS from the left ventricle. After removing the atria, the ventricles were digested in PBS containing 500 μg/mL of collagenase type II (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan), 200 μg/mL of CaCl2 (Nacalai Tesque, Kyoto, Japan), 0.05% trypsin (Sigma-Aldrich), and 10% FBS at 37 °C for 10 min with agitation. Digested samples were further dissociated by passing them through a 23-gauge needle three times. After treating with Red Blood Lysis buffer (BioLegend, CA, USA), isolated cells were filtered through a 40 μm strainer.
3
Surface Modification and Characterization of Titanium Alloys
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A commercially pure, medical-grade Ti plate (ISO5832-2) (Ti > 99.5%) was provided by Nilaco Co., Tokyo, Japan. Ti-6Al-4V alloy plate (Ti = balance, Al = 6.18, V = 4.27 mass%) and Ti-15Zr-4Nb-4Ta alloy plate (Ti = balance, Zr = 14.51, Nb = 3.83, Ta = 3.94, Pd = 0.16, and O = 0.25 mass%) were supplied by Kobelco Research Institute, Inc., Hyogo, Japan.
The chemical reagents (NaOH, CaCl2, ICl3, ICl, NaI and povidone iodine (PVP-I)) used for surface treatment were reagent-grade and purchased from Kanto Chemical Co., Inc., Tokyo, Japan. Reagent-grade NaCl, NaHCO3, KCl, K2HPO4·3H2O, MgCl2·6H2O, CaCl2, Na2SO4, tris-hydroxymethylaminomethane (CH2OH)3CNH2, and 1 M HCl were purchased from Nacalai Tesque, Inc., Kyoto, Japan, and used for the preparation of simulated body fluid (SBF). Minimum essential medium (MEM) was obtained from Gibco, Thermo Fisher Scientific, Waltham, MA, USA, and used for the cell culture test. Nutrient broth (Eiken Chemical Co., Ltd. Tochigi, Japan) and RPMI 1640 (Fujifilm Wako Pure Chemical Corp., Osaka, Japan) were used for the antibacterial activity test.
The chemical reagents (NaOH, CaCl2, ICl3, ICl, NaI and povidone iodine (PVP-I)) used for surface treatment were reagent-grade and purchased from Kanto Chemical Co., Inc., Tokyo, Japan. Reagent-grade NaCl, NaHCO3, KCl, K2HPO4·3H2O, MgCl2·6H2O, CaCl2, Na2SO4, tris-hydroxymethylaminomethane (CH2OH)3CNH2, and 1 M HCl were purchased from Nacalai Tesque, Inc., Kyoto, Japan, and used for the preparation of simulated body fluid (SBF). Minimum essential medium (MEM) was obtained from Gibco, Thermo Fisher Scientific, Waltham, MA, USA, and used for the cell culture test. Nutrient broth (Eiken Chemical Co., Ltd. Tochigi, Japan) and RPMI 1640 (Fujifilm Wako Pure Chemical Corp., Osaka, Japan) were used for the antibacterial activity test.
4
Embryonic Stem Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following reagents were obtained from the indicated manufacturers: bFGF (Wako), 4% paraformaldehyde (Wako), crystal violet (Wako), ES medium (ReproCELL), mTeSR1 (STEMCELL TECHNOLOGIES), trypsin (Life technologies), collagenase IV (Life technologies), KSR (Life technologies), TrypLE Express (Life technologies), Stempro34 SFM (Life technologies), DiI-Ac-LDL (Life technologies), Endothelial Serum-Free Medium (Life technologies), Laemmli buffer (Bio-Rad), LM411 (Biolamina), LM511 (Biolamina), Matrigel (BD), GFR-Matrigel (BD), Type IV collagen (BD), Cytofix (BD), Perm/Wash (BD), Fibronectin (Millipore), LM511-E8 fragment (Nippi), BMP4 (R&D systems), VEGF (R&D systems), RNeasy kit (QIAGEN), bovine serum albumin (BSA, sigma), DMEM/F12 (sigma), sodium dodecyl sulfate (SDS, Nacalai tesque) and CaCl2 (Nacalai tesque).
5
In Vitro Evaluation of Bioactive Polymer Screw HA Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The hydroxyapatite (HA)-forming ability of bioactive PSs was examined by incubation in simulated body fluid (SBF) as previously reported[26 (link)]. Briefly, samples were soaked in 24 ml of SBF at 36.5°C with the following ion concentrations approximating those of human blood plasma: Na+, 142.0; K+, 5.0; Ca2+, 2.5; Mg2+, 1.5; Cl–, 147.8; HCO3–, 4.2; HPO42-, 1.0; and SO42-, 0.5 mM. The SBF was prepared by dissolving reagent grade NaCl, NaHCO3, KCl, K2HPO4·3H2O, MgCl·6H2O, CaCl2, and Na2SO4 (Nacalai Tesque Inc., Kyoto, Japan) in ultrapure water, buffered at pH = 7.4 with tris (hydroxymethyl) aminomethane (CH2OH)3CNH2 and 1 M HCl (Nacalai Tesque Inc., Kyoto, Japan) at 36.5°C[27 (link)]. After soaking in SBF for three days, the samples were gently rinsed with ultrapure water and dried. HA formation on the surface of screws was evaluated by FE-SEM and energy dispersive X-ray analysis (EDX; EMAX-7000, Horiba Ltd., Kyoto, Japan). HA-forming ability was also evaluated on bioactive PSs that had been removed after insertion into the pedicle of cryopreserved canine lumbar spines.
6
Apatite Formation on Porous Titanium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cylindrical specimens of the porous Ti metals subjected to the various treatments given in Table 1 were broken and soaked in 30 mL of a simulated body fluid (SBF) having ion concentrations close to those of human blood plasma (Na+ = 142.0, K+ = 5.0, Mg2+ = 1.5, Ca2+ = 2.5, Cl– = 147.8, HCO3– = 4.2, HPO42– = 1.0, and SO42– = 0.5 mM) [19] (link), [20] (link) at 36.5°C. The SBF was prepared by dissolving reagent-grade NaCl, NaHCO3, KCl, K2HPO4·3 H2O, MgCl2.6 H2O, CaCl2, and Na2SO4 (Nacalai Tesque Inc., Japan). After 1 day, the specimens were removed from the SBF solution, gently washed with ultrapure water and dried in an oven at 40°C. Formation of apatite on their surfaces was examined by SEM and TF-XRD analysis.
7
3D Cell Culture in Calcium-Depleted FAD Medium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FAD medium (Biochrom GmbH, Berlin, Germany) consisted of Dulbecco’s modified Eagle’s medium (DMEM)/HAM’s F12 (3.5:1.1), 50 μM calcium chloride (CaCl2) and 4.5 g/L -glucose and was supplemented with 2.5% Chelex 100-treated fetal bovine serum (FBS), 0.18 mM adenine (Sigma-Aldrich), 0.5 μg/ml hydrocortisone (Sigma-Aldrich), 5 μg/ml insulin (Life Technologies, Carlsbad, CA, USA), 10–10 M cholera toxin (Sigma-Aldrich), 10 ng/ml epidermal growth factor (Sigma-Aldrich), 2 mM l -glutamine (Nacalai Tesque, Kyoto, Japan) and 1 mM sodium pyruvate (Wako, Osaka, Japan). Hereafter, this culture is referred to as the ‘complete FAD (c-FAD)’ medium. Serum calcium was removed by treating 500 ml FBS (HyClone, South Logan, UT, USA) with 20 g of Chelex 100 (Bio-Rad Laboratories, Hercules, CA, USA) [32 (link)]. For 3D cell cultures [33 (link)], the cells were cultured in the c-FAD medium supplemented with 1.2 mM CaCl2 (Nacalai Tesque), 10 ng/ml human keratinocyte growth factor (PeproTech, Rocky Hill, NJ, USA) and 0.283 mM l -ascorbic acid phosphate magnesium salt n-hydrate (Wako, Osaka, Japan), a stable derivative of ascorbic acid. Hereafter, this is referred to as the ‘FAD-3D medium’.
8
Simulated Body Fluid Soaking Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The samples subjected to the MA-heat or NaOH-HCl-heat treatments were soaked in 24 mL of a simulated body fluid (SBF) with various ion concentrations (Na+ 142.0, K+ 5.0, Ca2+ 2.5, Mg2+ 1.5, Cl− 147.8, HCO3− 4.2, HPO42− 1.0, and SO42− 0.5 mM) closed to human blood plasma at 36.5 °C. The SBF was prepared by dissolving reagent grade NaCl, NaHCO3, KCl, K2HPO4•3H2O, MgCl2•6H2O, CaCl2, and Na2SO4 (Nacalai Tesque Inc., Kyoto, Japan) in ultrapure water, and buffered at pH = 7.4 with tris(hydroxymethyl)aminomethane (CH2OH)3CNH2 and 1 M HCl (Nacalai Tesque Inc., Kyoto, Japan) at 36.5 °C [8 (link)]. After soaking in the SBF for 3 and 7 days, the samples were gently rinsed with ultrapure water, and dried.
9
Propagation of Spontaneous Ca2+ Transients in iCx26GJCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The propagation of spontaneous Ca2+ transients via gap junctions in iCx26GJCs was carried out using methods modified from a previous study (Schutz et al., 2010 (link)). Samples were incubated for 20 min at 37°C in high-calcium Dulbecco’s PBS (DPBS) containing 2 mM CaCl2 (Nacalai Tesque) in DPBS (Gibco) supplemented with 5 μM fluo-4 AM (Dojindo), 0.01% (w/v) pluronic F-127 (Invitrogen), and 250 μM sulfinpyrazone (Sigma) as loading medium. To record spontaneous Ca2+ transients, we replaced the loading medium with low-calcium DPBS, which contained 20 μM CaCl2.
The pharmacological study of iCx26GJCs was carried out using methods modified from previous studies (Schutz et al., 2010 (link), Tritsch et al., 2007 (link)). The spontaneous Ca2+ activity was recorded for 3 min, then the cells were superfused with PPADS (50 μM; Tocris) or FFA (50 μM; Sigma) for 5 min, followed by washout of the drug and superfusion with low-Ca2+ DPBS for 8 min. Sequential fluorescence images were acquired using a Nipkow disc confocal system (CSU22, Yokogawa) and AQUACOSMOS software (Hamamatsu Photonics). Live-cell imaging experiments were performed at near-physiological temperature (32°–35°C) or room temperature (24°–26°C). Signals were measured as relative changes of fluorescence intensity (Δf/f0), where f0 is the minimum fluorescence and f is the recorded fluorescence, and Δf = f − f0.
The pharmacological study of iCx26GJCs was carried out using methods modified from previous studies (Schutz et al., 2010 (link), Tritsch et al., 2007 (link)). The spontaneous Ca2+ activity was recorded for 3 min, then the cells were superfused with PPADS (50 μM; Tocris) or FFA (50 μM; Sigma) for 5 min, followed by washout of the drug and superfusion with low-Ca2+ DPBS for 8 min. Sequential fluorescence images were acquired using a Nipkow disc confocal system (CSU22, Yokogawa) and AQUACOSMOS software (Hamamatsu Photonics). Live-cell imaging experiments were performed at near-physiological temperature (32°–35°C) or room temperature (24°–26°C). Signals were measured as relative changes of fluorescence intensity (Δf/f0), where f0 is the minimum fluorescence and f is the recorded fluorescence, and Δf = f − f0.
10
Chitin Nanofiber Dispersion Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A chitin nanofiber/water dispersion (2 wt%, SFo-20002) was supplied by Sugino Machine, Ltd (Namerikawa, Japan). CaCl2 (>95.0% purity) and ethanol (>99.5% purity) were obtained from Nacalai Tesque, Inc. (Kyoto, Japan).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!