NRCM were cultured on 96-well plates (Sarstedt). The cells were transduced with shEGFP, shTiam or RGS3L–N460A encoding adenoviruses, and treated with FGF2 or PTX as described above. Thereafter, the cells were stimulated with 1 mM carbachol or solvent for 5 min at 37 °C. Myocytes were then loaded for 30 min with 5 µM 5(6)-carboxy-2′,7′-dichlorofluorescein (DCF-DA, Sigma-Aldrich) at 37 °C. After washing the cells with HEPES buffered salt solution (HBSS; 25 mM HEPES pH 7,4, 120 mM NaCl, 5,4 mM KCl, 1,8 mM CaCl2, 25 mM NaHCO3, 15 mM Glucose), DCF fluorescence was measured using the Envision 2102 Multilabel Reader with a set of FITC filters (excitation 485 ± 10 nM and emission at 535 ± 20 nM, PerkinElmer).
5 6 carboxy 2 7 dichlorofluorescein
Manufactured by Merck Group
Sourced in Germany
5(6)-carboxy-2′,7′-dichlorofluorescein is a chemical compound used as a fluorescent dye in various laboratory applications. It exhibits bright green fluorescence upon excitation with appropriate wavelengths of light. The compound is commonly used as a labeling agent in biochemical and cell biology research.
Automatically generated - may contain errors
Lab products found in correlation
Envision 2102 multilabel reader, by PerkinElmer (1 mentions)
96 well plate, by Sarstedt (1 mentions)
Losartan, by Merck Group (1 mentions)
Benzbromarone, by Merck Group (1 mentions)
Silicon oil, by Merck Group (1 mentions)
Glibenclamide, by Merck Group (1 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (1 mentions)
Paraffin oil, by Merck Group (1 mentions)
L proline, by Merck Group (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
Nahco3, by Chem-Lab (1 mentions)
Cacl2, by Chem-Lab (1 mentions)
2 protocols using 5 6 carboxy 2 7 dichlorofluorescein
1
Carbachol-Induced ROS Measurement
2
Comprehensive Reagents for Biochemical Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Vincristine-sulfate was acquired from LC chemicals (Woburn, Massachusetts); vinblastine (1 mg/ml) as well as methotrexate (2.5 mg/ml) were obtained from TEVA Pharma Belgium (Wilrijk, Belgium). Atazanavir-sulfate was obtained through the NIH Aids Reagent Program.
Benzbromarone, bromosulphophthalein, 5(6)-Carboxy-2′,7′-dichlorofluorescein (CDCF), digoxine, glibenclamide, losartan, paclitaxel, paraffin oil, L-proline, rifampycine, rifamycine SV and silicon oil (AP150) were purchased from Sigma-Aldrich (Schnelldorf, Germany). Dulbecco's modified eagle medium (DMEM; 1g/L glucose, 25 mM HEPES), fetal bovine serum (heatinactivated) (FBS), L-glutamine (200 mM), Hanks' Balanced Salt Solution (HBSS), penicillinstreptomycin mixture (10,000 IU potassium penicillin and 10,000 IU streptomycin sulfate per ml in 0.85 % saline), phosphate buffered saline (PBS), trypsine-EDTA mixture (170 U/ml and 200 mg/L, respectively) and Trypan blue stain (0.4 %) were obtained from Westburg (Leusden, The Netherlands). Dimethylsulfoxide (DMSO), methanol (MeOH) and Na-pyruvate were purchased from Acros Organics (Geel, Belgium). CaCl2 and NaHCO3 were obtained from Chem-lab NV (Zedelgem, Belgium) and NaCl from Fisher Chemical (Landsmeer, The Netherlands).
Benzbromarone, bromosulphophthalein, 5(6)-Carboxy-2′,7′-dichlorofluorescein (CDCF), digoxine, glibenclamide, losartan, paclitaxel, paraffin oil, L-proline, rifampycine, rifamycine SV and silicon oil (AP150) were purchased from Sigma-Aldrich (Schnelldorf, Germany). Dulbecco's modified eagle medium (DMEM; 1g/L glucose, 25 mM HEPES), fetal bovine serum (heatinactivated) (FBS), L-glutamine (200 mM), Hanks' Balanced Salt Solution (HBSS), penicillinstreptomycin mixture (10,000 IU potassium penicillin and 10,000 IU streptomycin sulfate per ml in 0.85 % saline), phosphate buffered saline (PBS), trypsine-EDTA mixture (170 U/ml and 200 mg/L, respectively) and Trypan blue stain (0.4 %) were obtained from Westburg (Leusden, The Netherlands). Dimethylsulfoxide (DMSO), methanol (MeOH) and Na-pyruvate were purchased from Acros Organics (Geel, Belgium). CaCl2 and NaHCO3 were obtained from Chem-lab NV (Zedelgem, Belgium) and NaCl from Fisher Chemical (Landsmeer, The Netherlands).
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