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Cacl2

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Calcium chloride (CaCl2) is a chemical compound that is commonly used in laboratory settings. It is a white, crystalline solid that is soluble in water and has a high melting point. CaCl2 is known for its ability to absorb moisture from the air, making it a useful desiccant. It can also be used as an electrolyte and a source of calcium ions in various chemical reactions and applications.

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Lab products found in correlation

Sodium chloride (nacl), by Avantor (14 mentions) Potassium chloride (kcl), by Avantor (11 mentions) Mgso4, by Avantor (7 mentions) Glucose, by Avantor (5 mentions) Hepes, by Merck Group (4 mentions) Kh2po4, by Avantor (4 mentions) Mgcl2, by Avantor (4 mentions) Tween 20, by Merck Group (3 mentions) Bovine serum albumin (bsa), by Merck Group (3 mentions) Acetylcholine chloride, by Merck Group (2 mentions) Bovine serum albumin (bsa), by Santa Cruz Biotechnology (2 mentions) Sodium nitrite, by Merck Group (2 mentions) Kh2po2, by Avantor (2 mentions) Phenylephrine, by Merck Group (2 mentions) Sodium nitroprusside, by Merck Group (2 mentions) Rpmi 1640 medium, by Cambrex (2 mentions) Sodium chloride (nacl), by Merck Group (2 mentions) Gallic acid, by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Carl Roth (2 mentions) Sodium hydroxide (naoh), by Avantor (2 mentions)

43 protocols using cacl2

1

Nuclei Isolation for Single-Cell RNA-Seq

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Fresh-frozen tissues were disaggregated in 1 mL of custom
nuclear extraction buffer (see Table S1 for all
combinations used) with mild chopping by Tungsten Carbide Straight 11.5
cm Fine Scissors (14558-11, Fine Science Tools, Foster City, CA) for 10
minutes on ice. Large debris were removed with a 40 μm strainer
(Falcon). An additional 1 mL of buffer was used to wash the filter
before proceeding to fluorescence-activated cell sorting (FACS). For
droplet-based RNA-Seq, nuclei were isolated as described above, but with
the addition of 3 ml of ST (Salts and Tris; Table S1) to extracted
nuclei. Nuclei were then pelleted at 500g for 5 mins at 4°C.
Supernatant was discarded and the nuclei pellet was resuspended in
100-500 μL of ST buffer (Salts and Tris; Table S1) before filtering
through a 40 μm strainer-capped round bottom tube (Falcon).
For the ENS atlas, the CST (“RAISIN”) composition
was: 0.49% (w/v) CHAPS (Cat#220201-1GM, EMD Millipore), 146 mM NaCl
(Cat#S6546-1L, Sigma-Aldrich), 1mM CaCl2 (Cat#97062-820, VWR), 21 mM
MgCl2 (Cat#M1028-10X1ML, Sigma-Aldrich), 10 mM Tris pH 8.0 (CAT#AM9855G,
Thermo Fisher Scientific). The TST (“INNER Cell”)
composition was: 146 mM NaCl (Cat#S6546-1L, Sigma Aldrich), 1 mM CaCl2
(Cat#97062-820, VWR), 21 mM MgCl2 (Cat#M1028-10X1ML, Sigma Aldrich), 10
mM Tris pH 8.0 (CAT#AM9855G, Thermo Fisher Scientific).
Drokhlyansky E., Smillie C.S., Van Wittenberghe N., Ericsson M., Griffin G.K., Eraslan G., Dionne D., Cuoco M.S., Goder-Reiser M.N., Sharova T., Kuksenko O., Aguirre A.J., Boland G.M., Graham D., Rozenblatt-Rosen O., Xavier R.J, & Regev A. (2020). The human and mouse enteric nervous system at single cell resolution. Cell, 182(6), 1606-1622.e23.
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2

3D-Printed Scaffold for Breast Cancer Cell Culture

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For the 3D‐printed scaffold (3DPS) synthesis, alginate of 8% (wt/vol %; Protanal LF 10/60; FMC) and 5% hydroxyapatite (wt/vol %; Sigma‐Aldrich) were mixed in water (Synergy Elix 15; Merck) using an Ultra‐Thurrax T50 digital dispenser (IKA), equipped with an S 25N‐25G dispensing tool, at 5000 rpm overnight and printed in four layers (⌀20 × 2 mm; grid distance 1.5 mm, 90°) using a bioplotter (Envisiontech) equipped with 400 µm extrusion needles. Each layer was cross‐linked in 0.1 M CaCl2 (VWR) and ready prints were stored in 0.1 M CaCl2 (VWR) at 4°C for up to 2 weeks.
For cell culture, 3DPS were washed in cell culture media, placed in 24 wells‐plates and seeded with 3 × 105 MCF7 cells in supplemented DMEM. After 24 h, 3DPS were moved to a six well‐plates with fresh media, and this process was repeated every 4 days to a total culturing time of 21 days. After that, 3DPS were treated with 5‐FU and DOX at the 10X concentration as the PDS treatments. For RNA extraction, 3DPS were washed twice in cell media, lysed in 700 µl QIAzol (Qiagen) and homogenized for 2 × 2.5 min at 25 Hz. Automated isolation of total RNA from lysates was performed in a QIAcube machine (Qiagen) using a RNeasy Micro Kit (Qiagen) with QIAzol extraction directives. Reverse transcription and qPCR were performed as explained above.
Leiva M.C., Garre E., Gustafsson A., Svanström A., Bogestål Y., Håkansson J., Ståhlberg A, & Landberg G. (2020). Breast cancer patient‐derived scaffolds as a tool to monitor chemotherapy responses in human tumor microenvironments. Journal of Cellular Physiology, 236(6), 4709-4724.
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3

Planar lipid bilayer formation for KirBac3.1 channel

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KirBac3.1 proteoliposomes were incorporated into planar lipid bilayers separating two aqueous chambers (cis and trans). Bilayers were produced using the Mueller method52 (link). Bilayers were formed across an aperture in a delrin cup with diameter 150–250 μm using a lipid mixture of phosphatidylethanolamine, phosphatidylcholine and phosphatidylserine (7:2:1 wt/wt, Avanti Polar Lipids) in n-decane (50 mg ml−1, ICN Biomedicals). The cis chamber contained 250 mM KCl and 1.0 mM CaCl2 and the trans chamber contained 50 mM KCl and 0.1 mM CaCl2. Proteoliposomal fusion with bilayers was initiated by adding 5 μl aliquots of the proteoliposome preparation (described above) to the cis bath whilst stirring. During experiments, the composition of the cis solution was altered by a perfusion system53 (link) that allowed exposure of single channels to additions of spermine or KCl/NaCl substitutions within ~1 s. All solutions were pH buffered using 10 mM N-tris[hydroxymethyl] methyl-2-aminoethanesulfonic acid (TES; ICN Biomedicals) and titrated to pH 7.4 using KOH (ICN Biomedicals). KCl was obtained from Aldrich and CaCl2 from BDH Chemicals. Experiments were carried out at room temperature (23 ± 2 °C).
Jin R., He S., Black K.A., Clarke O.B., Wu D., Bolla J.R., Johnson P., Periasamy A., Wardak A., Czabotar P., Colman P.M., Robinson C.V., Laver D., Smith B.J, & Gulbis J.M. (2022). Ion currents through Kir potassium channels are gated by anionic lipids. Nature Communications, 13, 490.
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4

Vascular Reactivity Pharmacological Agents

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Phenylephrine, acetylcholine chloride, sodium nitrite, sodium nitroprusside, L-NAME and Tween-20 were purchased from Sigma Chemicals (St Louis, MO, USA). NaCl was purchased from Calbiochem® Merck (Darmstadt, Germany). MgSO4, KCl, KH2PO2, glucose and CaCl2 were purchased from BDH Laboratory Supplies (Poole, UK). Bovine serum albumin (BSA) was purchased from Santa Cruz (Dallas, Texas, USA). All compounds were dissolved in deionized water. The concentrations stated are expressed as final molar concentrations in the buffer. At the concentrations used, the vehicles had no significant effect on vascular responses (data not shown).
Ling W.C., Murugan D.D., Lau Y.S., Vanhoutte P.M, & Mustafa M.R. (2016). Sodium nitrite exerts an antihypertensive effect and improves endothelial function through activation of eNOS in the SHR. Scientific Reports, 6, 33048.
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5

Preparation of Pharmacological Reagents

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Phenylephrine, acetylcholine chloride, sodium nitrite, sodium nitroprusside, and Tween-20 were purchased from Sigma Chemicals (St Louis, MO, USA). Angiotensin II was purchased from Sigma Chemicals (USA). EGCG was purchased from Cayman. NaCl was purchased from Calbiochem® Merck (Darmstadt, Germany). MgSO4, KCl, KH2PO2, glucose and CaCl2 were purchased from BDH Laboratory Supplies (Poole, UK). Bovine serum albumin (BSA) was purchased from Santa Cruz (Dallas, Texas, USA). All compounds were dissolved in deionized water. The concentrations stated are expressed as final molar concentrations in the buffer.
Mohd Sabri N.A., Lee S.K., Murugan D.D, & Ling W.C. (2022). Epigallocatechin gallate (EGCG) alleviates vascular dysfunction in angiotensin II-infused hypertensive mice by modulating oxidative stress and eNOS. Scientific Reports, 12, 17633.
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6

Isolation of Human Neutrophils

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Venous blood from all participants was collected in 30 mL syringes (containing 5 mL acid citrate dextrose for 25 mL whole blood). Neutrophils were isolated using the Ficoll-Paque gradient method, as previously described [67 (link), 68 (link)]. Upon isolation, neutrophils were resuspended in phenol-free RPMI-1640 medium (Cambrex Bio Science, Walkersville, MD) supplemented with (1) 25 mM HEPES (N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid) (Sigma-Aldrich, Oakville, ON, Canada), (2) 1% penicillin/streptomycin/ Glutamax (VWR Intl., Montreal, QC, Canada), (3) 1 mM CaCl2 (BDH Chemicals, Toronto, ON, Canada) and (4) 5% FBS (Fetal Bovine serum; VWR) (termed complete RPMI). Contamination by PMBCs was less than 0.1% as determined by morphological analysis and flow cytometry. Cell viability of neutrophils were greater than 98%, as assessed by Trypan blue dye exclusion assay.
Charles E., Dumont B.L., Bonneau S., Neagoe P.E., Villeneuve L., Räkel A., White M, & Sirois M.G. (2021). Angiopoietin 1 release from human neutrophils is independent from neutrophil extracellular traps (NETs). BMC Immunology, 22, 51.
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7

Evaluation of Antioxidant and Vasoactive Compounds

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ISO, hexadecyltrimethylammonium, trichloroacetic acid, carbachol, butylated hydroxytoluene, D-glucose, gallic acid, quercetin, thiobarbituric acid, and noradrenaline were purchased from Sigma-Aldrich (Germany). Propranolol was purchased from Teva Santé (France). DMSO, EDTA, NADH, Folin–Ciocalteu reagent, and sodium pyruvate were from Carl-Roth (Kalshur, Germany). L-NAME and glibenclamide were bought from Enzo Life Sciences (Lausen, Switzerland). NaHCO3 was purchased from Riedel‐de Haën AG. KH2PO4, KCl, CaCl2, MgSO4, NaCl, and orthophosphoric acid were bought from BDH (chemicals Ltd Poole England).
Nguelefack-Mbuyo E.P., Nokam F., Tchinda N.L., Goumtsa A.F., Tsabang N, & Nguelefack T.B. (2022). Vasorelaxant and Antioxidant Effects of Aframomum pruinosum Gagnep. (Zingiberaceae) Seed Extracts May Mediate Their Cardioprotective Activity against Isoproterenol-Induced Myocardial Infarction. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 7257448.
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8

Pharmacological Modulation of Adenosine A2A Receptors

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NaCl, NaOH, NaH2PO4, NaHCO3, KCl, BaCl2, CaCl2, MgCl2, sucrose, CsCl, NMDG and glucose were purchased from BDH Laboratory Supplies (Poole, England). HEPES, EGTA, ATP and GTP, were purchased from Sigma (St. Louis, MO). Tetrodotoxin (TTX) was purchased from Ascent Scientific (Bristol, UK) or Alomone Labs (Jerusalem, Israel). The concentrations of the A2A receptor agonist CGS21680 (1 μM) and antagonist SCH58261 (10 μM) were chosen on the basis of ligand‐binding studies from Cunha et al. (1999 (link)) and from Zocchi et al. (1996 (link)), respectively.
Coppi E, & Gibb A.J. (2022). Selective block of adenosine A2A receptors prevents ischaemic‐like effects induced by oxygen and glucose deprivation in rat medium spiny neurons. British Journal of Pharmacology, 179(20), 4844-4856.
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9

Isolation and Characterization of Neutrophils

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Venous blood from all participants was collected in serum-separating tubes (SST) (3.5-mL blood volume) and in 30 mL syringes (containing 5 mL of acid citrate dextrose for 25 mL of whole blood). The SST tubes were centrifuged to obtain serum, which was aliquoted and frozen at −80 °C. Neutrophils were isolated using the Ficoll–Paque gradient method, as previously described [28 (link)]. Upon isolation, neutrophils were resuspended in phenol-free RPMI-1640 medium (Cambrex Bio Science, Walkersville, MD, USA) supplemented with 25 mM HEPES (N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid) (Sigma–Aldrich, Oakville, ON, Canada), 1% penicillin/streptomycin/Glutamax (VWR Intl., Montreal, QC, Canada), 1 mM CaCl2 (BDH Chemicals, Toronto, ON, Canada), and 5% FBS (Fetal Bovine serum; VWR Intl., Montreal, QC, Canada) (termed complete RPMI). Contamination by PMBCs was less than 0.1% as determined by morphological analysis and flow cytometry. Cell viability of neutrophils was greater than 98%, as assessed by Trypan blue dye exclusion assay.
Dumont B.L., Neagoe P.E., Charles E., Villeneuve L., Tardif J.C., Räkel A., White M, & Sirois M.G. (2024). Low-Density Neutrophils Contribute to Subclinical Inflammation in Patients with Type 2 Diabetes. International Journal of Molecular Sciences, 25(3), 1674.
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10

Tardigrade Tun Formation and Survival

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Tardigrades (30 animals per replicate, 4 replicates per concentration/condition) were transferred in minimal media (<100 μL) on 35-mm plastic Petri dishes and dosed with working concentrations of 50, 75, 100, and 150 mM CaCl2 (Amresco, Dallas, TX) or 225, 300, 450, or 600 mM sucrose (Millipore, Burlington, MA). Tardigrades were manually counted every half hour for tun formation; counting stopped once all conditions were 100% in tuns (12 h for CaCl2 and 6 h for sucrose) for 5 and 1 h, respectively, and tun formation was monitored. Animals were transferred to new plates containing culturing media and monitored for recovery. Survival for each concentration was defined as tardigrades exhibiting controlled movement of limbs and/or body post-recovery treatment. Surviving tardigrades were counted after 24 h.
Smythers A.L., Joseph K.M., O’Dell H.M., Clark T.A., Crislip J.R., Flinn B.B., Daughtridge M.H., Stair E.R., Mubarek S.N., Lewis H.C., Salas A.A., Hnilica M.E., Kolling D.R, & Hicks L.M. (2024). Chemobiosis reveals tardigrade tun formation is dependent on reversible cysteine oxidation. PLOS ONE, 19(1), e0295062.
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