The cardiac function of mice was measured at baseline (day 0) before the beginning of the experiment. Animals were then randomly assigned to DOX or vehicle treatment group. After baseline cardiac assessment, mice in the DOX treatment group received a single intraperitoneal injection of DOX (20 mg/kg; MedacGmbH, Wedel, Germany) in max 150 µL NaCl (B. Braun Melsungen AG, Melsungen, Germany), whereas mice in the vehicle treatment group received a single injection of 150 µL NaCl only. Following published conversions, this DOX dose equals ~55 mg/m2 [45 (link),46 ]. Mice were weighed and visually investigated every day to check for worsening overall fitness. On day 6 after injection, mice were again measured for cardiac function. Afterward, animals were killed by exsanguination in deep isoflurane (Baxter GmbH, Unterschleißheim, Germany) narcosis after injection of Ketamine/Xylazine (100/20 mg/kg; Hameln Pharma, Hameln, Germany/Bela Pharm, Vechta, Germany). The heart was extracted after perfusing the animal free of blood with PBS without calcium or magnesium (PBS−Ca/−Mg; ThermoFisher Scientific, Waltham, MA, USA) including 20 IE/mL of heparin (Leo Pharma GmbH, Neu-Isenburg, Germany). From the extracted heart, a single midventricular slice (~2 mm thickness) was obtained for histology and fixated in 4% PFA at RT overnight, while the rest of the heart was used for flow cytometry.
Sodium chloride (nacl)
Manufactured by B. Braun
Sourced in Germany, Switzerland, United Kingdom, United States
NaCl is a laboratory product that serves as a source of sodium and chloride ions. It is a white, crystalline solid that is commonly used in various scientific and medical applications. NaCl is a fundamental chemical compound with a simple chemical formula and well-understood properties.
Automatically generated - may contain errors
Lab products found in correlation
Xylavet, by CP-Pharma (2 mentions)
Microlance 3, by BD (2 mentions)
Ketanest s, by Pfizer (2 mentions)
Granulocyte monocyte progenitor dendritic cell medium, by CellGenix (2 mentions)
Ficoll paque plus, by GE Healthcare (2 mentions)
Suprarenin, by Sanofi (2 mentions)
Clonidin, by Ratiopharm (2 mentions)
Polycal, by Nutricia (2 mentions)
Boso medicus prestige, by Bosch (2 mentions)
Phosphate buffered saline (pbs), by Merck Group (2 mentions)
Collagenase 2, by Worthington (1 mentions)
Mk 8825, by Merck Group (1 mentions)
Sodium iodide, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Peracetic acid, by Merck Group (1 mentions)
4 methylcatechol, by Merck Group (1 mentions)
Pertussis toxin ptx, by Merck Group (1 mentions)
53 protocols using sodium chloride (nacl)
1
Doxorubicin-Induced Cardiac Dysfunction in Mice
2
Cartilage Biopsy and Cryopreservation
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Cartilage biopsies were collected during septum or septum rhinoplasty operations at the Department of Oto-Rhino-Laryngology, Head and Neck Surgery at the University of Ulm. The biopsies were collected after the patients’ written consent had been obtained and the protocol was approved by the local ethics committee (University of Ulm, no.152/08). All patients were anonymized, although age and gender were recorded. The cells from a donor were only used in this experiment if there could enough cells be isolated to analyse them on each point of time.
The biopsies were transported to the laboratory in 0.9% sodium chloride (B. Braun, Melsungen, Germany). On arrival, each biopsy was cut into smaller pieces (approximately 1 × 1 mm); this was followed by digestion in standard culture medium Dulbecco’s modified Eagle’s medium and Ham’s F-12 nutrient mixture (DMEM/HAMS F-12; 10% standardized foetal bovine serum (FBS); 0.5% gentamicin) with 0.3% collagenase II (Worthington Biochemical Corporation, Lake Wood, USA) for 16 h at 37°C in a waterbath. The cells were then washed in standard culture medium, counted using a Neubauer counting chamber, cryopreserved for later use with freezing media (DMEM/HAMS F-12; 40% foetal calf serum (FCS); 20% dimethyl sulfoxide (DMSO)) and stored in liquid nitrogen.
The biopsies were transported to the laboratory in 0.9% sodium chloride (B. Braun, Melsungen, Germany). On arrival, each biopsy was cut into smaller pieces (approximately 1 × 1 mm); this was followed by digestion in standard culture medium Dulbecco’s modified Eagle’s medium and Ham’s F-12 nutrient mixture (DMEM/HAMS F-12; 10% standardized foetal bovine serum (FBS); 0.5% gentamicin) with 0.3% collagenase II (Worthington Biochemical Corporation, Lake Wood, USA) for 16 h at 37°C in a waterbath. The cells were then washed in standard culture medium, counted using a Neubauer counting chamber, cryopreserved for later use with freezing media (DMEM/HAMS F-12; 40% foetal calf serum (FCS); 20% dimethyl sulfoxide (DMSO)) and stored in liquid nitrogen.
3
Ovariectomized Mice CGRP Antagonist
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The ovariectomized mice were randomly allocated to treatment with vehicle (saline) or the CGRP-antagonist MK-8825 (Merck, United Kingdom). Twenty minutes before running, 200 μl of sodium chloride (0.9%; B. Braun, Germany) or CGRP-antagonist MK-8825 at a dose of either 10 mg/kg body weight or 50 mg/kg body weight of (8750 μg/ml in 0.9% NaCl) was injected intraperitoneally.
4
Radiolabeling of (m)TMSBG with I-124
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[[3-(Trimethylsilyl)phenyl]methyl] guanidine sulphate (mTMSBG) was purchased from ABX advanced biochemical compounds (Dresden, Germany). Sodium iodide, trifluoroacetic acid, potassium phosphate (monobasic, Ph Eur grade), sodium bicarbonate, phosphoric acid, peracetic acid (32 % v/v in acetic acid) and ethanol were purchased from Sigma Aldrich Ltd (Gillingham, UK). 0.9 % sodium chloride was purchased from B Braun (Sheffield, UK). 124I as [124I]Sodium iodide (t1/2 = 4.18 days, β+ = 25.6 %) was supplied by Perkin Elmer and produced by BV Cyclotron VU (Amsterdam, The Netherlands) by a 124Te(p,n)124I reaction on enriched tellurium (124Te). It was isolated by dry distillation followed by chemical trapping in 0.2 M sodium hydroxide (~1.1 GBq ml−1).
5
Dexmedetomidine Injection in Neonatal Pups
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The pups were injected at P7 with 25 µg/kg of dexmedetomidine intraperitoneally (IP) using a 0.1 mL volume, whereas the control animals received a corresponding volume of saline (sodium chloride 0.9% B Braun, Mississauga, ON, Canada). The animal’s body temperature was maintained by a heated blanket and nitrile gloves filled with warm water, which was replaced every 20 min. The respiratory status of the animal was monitored by counting the number of breaths per minute every 10 min after the first injection for 30 min, to ensure the health of the pup. Similarly, oxygen saturation (SpO2) was monitored (Nonin Medical, Minneapolis, MN, USA) every 30 min after the first injection for a period of 1 h to further ensure the pup’s health.
6
Platelet Aggregation Induced by Collagen, AA, and TRAP
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A total of 300 µL of whole blood was diluted with the same volume of 0.9 % sodium chloride (B.Braun, Melsungen, Germany) preheated to 37 °C and incubated with 5 µL of a tested compound dissolved in DMSO (Penta, Prague, Czech Republic) at a final concentration of 0.8% for 3 min at 37 °C. Platelet aggregation was then induced by addition of collagen (Diagnostica a.s., Czech Republic), arachidonic acid (AA) or Thrombin Receptor Activating Peptide (TRAP) (Roche Holding AG, Basel, Switzerland) and monitored for 6 min. The dose of inducer was initially set to the minimal concentration which caused maximal aggregation. The second calibration was carried out by use of standards, 4-methylcatechol in the case of AA- and collagen-triggered aggregation (Sigma-Aldrich) or vorapaxar for thrombin aggregation (Selleck Chemicals GmbH, Munich, Germany). The final concentrations of inducers were in the range of 0.16–2.42 µg/mL for collagen, 25–196 µM for AA and 0.8–9.8 µM for TRAP. For comparison, also the clinically used standard ASA was employed.
7
Experimental Autoimmune Optic Neuritis Induction
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WT (WT ONA) and KO (KO ONA) mice were immunized intraperitoneally with ONA (1 mg/ml) mixed with incomplete Freund's adjuvants (FA; 50 μl) and 1 μg pertussis toxin (PTx; both Sigma Aldrich, St. Louis, MO, USA) as described (49 (link)). To generate the ONA homogenate, fresh bovine eyes were obtained from a local slaughterhouse (Schlachthaus Wuppertal, Germany). As previously described, optic nerves were cut off behind the optic nerve head, cleaned from surrounding tissue and the dura mater was removed. Nerves were pulverized in a cooled mortar and then suspended in phosphate-buffered saline (PBS) (5 (link)). A final concentration of 1 mg/ml was set. FA acted as an immunostimulatory and PTx was given to ensure the permeability of the blood retina barrier. Intraperitoneal PTx-application was repeated 2 days after immunization. Booster injections containing half of the initial dose were given intraperitoneally 4 and 8 weeks after initial immunization. The control groups (WT CO; KO CO) were injected with 1 ml sodium chloride (B. Braun Melsungen AG, Melsungen, Germany), FA and PTx. Ten weeks after immunization, retinae, and optic nerves were explanted for immunohistochemistry, quantitative real time PCR (RT-qPCR), and Western blot analyses. For RT-qPCR and Western blot, retinal as well as optic nerve tissue of both eyes from one animal were pooled.
8
Experimental Autoimmune Retinopathy in Mice
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WT (WT ONA) and KO (KO ONA) mice were immunized with ONA (1 mg/ml) mixed with incomplete Freund`s adjuvants (FA) and 1 µg pertussis toxin (PTx; both Sigma Aldrich, St. Louis, MO, USA) according to the previously described pilot study (Reinehr et al., 2019) . FA acted as an immunostimulatory and PTx was given to ensure the permeability of the blood retina barrier. PTx-application was repeated 2 days after immunization. Booster injections containing half of the initial dose were given 4 and 8 weeks after initial immunization. The control groups (WT CO; KO CO) were injected with 1 ml sodium chloride (B. Braun Melsungen AG, Melsungen, Germany), FA and PTx. 10 weeks after immunization, retinae and optic nerves were explanted for immunohistochemistry, quantitative real time PCR (RT-qPCR), and Western blot analyses. For RT-qPCR and Western blot, we pooled retinal and optic nerve tissue of both eyes.
9
Injection Pressure Assessment for Testicular Procedures
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The injection pressure of the two methods used in the experiments was measured in four piglets injected using the two-step method and seven piglets injected using the one-step F method. Immediately after euthanasia injection with 0.6 mL 0.9% sodium chloride (B. Braun, Melsungen, Germany) was performed via the one-step F or two-step method, as described. Before injection, the system was flushed with 0.9% sodium chloride, and all air bubbles were removed from the hoses. Pressure was measured via a pressure sensor (Xtrans, Codan pvb Medical GmbH) and recorded every four seconds (EIM-B, EIM-A, HAEMODYN software, Hugo Sachs Elektronik—Harvard Apparatus GmbH, March-Hugstetten, Germany). Injections into the air were performed to determine the baseline system pressure due to the equipment. The mean maximum pressures of both injection techniques into the testicular tissue and air were calculated. To determine the duration of injection, the time from injection into the first testis to withdrawal of the cannula from the second testis was measured.
10
Cecal Ligation and Puncture in Mice
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For the investigation of post-septic immunological consequences a cecal ligation and puncture (CLP) mouse model was used as described before (21 (link)). Briefly, mice were anesthetized by an intraperitoneal injection of 100 mg/kg ketamine (Ketanest®S, Pfizer Pharma, Berlin, Germany) and 20 mg/kg xylazine (Xylavet, CP-Pharma, Burgdorf, Germany). After median laparotomy, cecum was mobilized, ligated (5 mm).and punctured once with a 23 G needle (BD Microlance™ 3, BD Medical, Heidelberg, Germany). Fecal content was gently extruded and the cecum afterwards relocated. Mice were supplemented with 400 μL 0.9% sodium chloride (B. Braun, Melsungen, Germany), given directly in the abdominal cavity and abdomen was closed thereafter with a double suture. Control animals received a sham surgery without cecal ligation and puncture. For pain relieve, mice were treated with 0.05 mg/kg bodyweight buprenorphine (Temgesic, RB Pharmaceuticals, Slough, UK) every 8 h for 2 days after surgery. In total, 15 animals received a CLP and 9 animals a sham surgery. All animals were subsequently housed till euthanasia 12 weeks after intervention (Figure 1A ).
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