Both the left and right NG neurons were quickly excised from cervical dislocation-killed rats. After desheathed in ice-cold sterile Krebs solution, the ganglia were incubated for 50 min in digestion buffer of collagenase type I (1 mg/mL, Sigma) and trypsin II-S (0.5 mg/mL, Sigma) in 5 mL DMEM medium (Gibco, Gaithersburg, MD, USA) at 37 °C. The enzymatic reaction was terminated by soybean trypsin inhibitor type II-S (0.625 mg/mL, Sigma). The ganglia were then transferred to 1 mL DMEM tissue culture medium containing 10% fetal bovine serum (Gibco). A single-cell suspension was created by repeated mechanical trituration using a plastic Pasteur pipe. The NG neurons were plated onto poly-l-lysine-coated dishes and then incubated at 37 °C in a 5% CO2 incubator (Thermo Forma, Hamilton, NJ, USA).
Soybean trypsin inhibitor type 2 s
Manufactured by Merck Group
Soybean trypsin inhibitor (type II-S) is a laboratory reagent used for the inhibition of trypsin, a serine protease enzyme. It is extracted from soybeans and functions by binding to and blocking the active site of trypsin, preventing its proteolytic activity.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase type ia, by Merck Group (4 mentions)
Trypsin type 2 s, by Merck Group (4 mentions)
Dnase type 4, by Merck Group (4 mentions)
Trypsin, by Merck Group (3 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Dmem tissue culture medium, by Thermo Fisher Scientific (1 mentions)
Trypsin 2 s, by Merck Group (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
5 protocols using soybean trypsin inhibitor type 2 s
1
Isolation and Culture of Rat Nodose Ganglia Neurons
2
Isolation and Culture of Rat Dorsal Root Ganglia
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All experimental protocols were approved by the animal research ethics committee of Hubei University of Science and Technology. Sprague-Dawley male rats (5–6 weeks old) were anesthetized and then killed. The DRGs were removed and chopped with thin spring scissors. The minced ganglia were transferred to a test tube containing Dulbecco’s modified Eagle’s medium (DMEM, Sigma) and incubated in a shaking for 25–30 min at 35°C. Incubation solution contained 1.0 mg/ml collagenase (type I-A, Sigma), 0.5 mg/ml Trypsin (type II-S, Sigma) and 0.1 mg/ml DNase (type IV, Sigma). Trypsin digestion was terminated by adding1.25 mg/ml Soybean Trypsin inhibitor (type II-S, Sigma). The cells were cultured in DMEM supplemented with 10% fetal bovine serum and 100 ng/ml never growth factor (NGF) for 12–24 h at 37°C in a water saturated atmosphere with 95% O2 and 5% CO2.
3
Rat DRG Isolation and Dissociation
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All experimental protocols were approved by the animal research ethics committee of Hubei University of Science and Technology. All procedures were performed to minimize the suffering of animals. Male Sprague-Dawley rats (5- to 6-week-old) were sacrificed. The DRGs were removed and minced with fine spring scissors. The ganglion fragments were placed in a flask containing 5 mL of Dulbecco's modified Eagle's medium (DMEM, Sigma). DMEM contained trypsin (type II-S, Sigma) 0.5 mg/mL, collagenase (type I-A, Sigma) 1.0 mg/mL and DNase (type IV, Sigma) 0.1 mg/mL, and were incubated at 35 °C in a shaking water bath for 25–30 min. Soybean trypsin inhibitor (type II-S, Sigma) 1.25 mg/mL was then added to stop trypsin digestion.
4
Isolation of Dorsal Root Ganglia
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All studies were designed to minimize the sufferings of animals and approved by the animal research ethics committee of Hubei University of Science and Technology (No. 2020–08). Sprague-Dawley male rats (5–6 weeks old) were anesthetized and then killed. The DRGs were removed and chopped with thin spring scissors. The minced ganglia were transferred to a test tube containing Dulbecco’s modified Eagle’s medium (DMEM, Sigma) and incubated in a shaking for 25–30 min at 35°C. Incubation solution contained 1.0 mg/mL collagenase (type I-A, Sigma), 0.5 mg/mL Trypsin (type II-S, Sigma), and 0.1 mg/mL DNase (type IV, Sigma). Trypsin digestion was terminated by adding 1.25 mg/mL Soybean Trypsin inhibitor (type II-S, Sigma).
5
Isolation and culturing of rat DRG neurons
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All experimental protocols were approved by the animal research ethics committee of Hubei University of Science and Technology (No. 2020-07). All procedures were made to minimize the sufferings of animals. Sprague-Dawley male rats (6- to 7-week-old) were anesthetized with 7% chloral hydrate. The DRGs were taken out and minced with fine spring scissors. The ganglion fragments were placed in a flask containing 5ml of Dulbecco’s modified Eagle’s medium (DMEM, Sigma). DMEM contained trypsin (type II-S, Sigma) 0.5 mg/ml, collagenase (type I-A, Sigma) 1.0 mg/ml, and DNase (type IV, Sigma) 0.1 mg/ml and was incubated at 35°C in a shaking water bath for 25–30 min. Soybean trypsin inhibitor (type II-S, Sigma) 1.25 mg/ml was then added to stop trypsin digestion. Freshly dissociated neurons were placed into a 35-mm Petri dish and kept for at least 1 h in normal external solution before the start of electrophysiological experiments.
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