The induction of apoptosis was monitored using the TUNEL method. The TUNEL assay was performed in accordance with manufacturer's protocol of the TUNEL apoptosis kit (R&D Systems, Inc.).
Tunel apoptosis kit
Manufactured by R&D Systems
Sourced in United States
The TUNEL apoptosis kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes the TUNEL (Terminal deoxynucleotidyl Transferase dUTP Nick End Labeling) assay, which labels the fragmented DNA that is a hallmark of apoptotic cells. This enables the visualization and analysis of apoptotic cells within a sample.
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3 protocols using tunel apoptosis kit
1
Apoptosis Detection via TUNEL Assay
2
Quantifying DRG Neuron Apoptosis
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DRG culture was quickly washed with PBS, and fixed by 4% paraformaldehyde (PFA, Sigma-Aldrich, USA) for 30 min. The apoptosis of DRG neurons was examined by a terminal deoxyribonucleotidyl transferase (TdT)-mediated biotin-16-dUTP nick-end labeling (TUNEL) Apoptosis Kit (R&D Systems, USA) according to the manufacturer’s protocol. A mouse monoclonal NeuN antibody (Millipore, USA) was used to identify the DRG neurons. Fluorescent images were then examined under an upright fluorescent microscope (BX51, Olympus, Japan). Relative apoptosis was quantified as the percentage of TUNEL-positive cells among all NeuN-positive cells for each experimental condition.
3
Measuring Apoptosis by Flow Cytometry and TUNEL
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Cells were washed twice with cold PBS, and then resuspended in 1× binding buffer (BD Biosciences, San Jose, CA, USA) at a concentration of 1×106 cells/mL. Then, 100 mL of the solution (1×105 cells) was transferred to a 5 mL culture tube and stained with 5 mL each of allophycocyanin–annexin V (BD Biosciences) and 50 mg/mL propidium iodide (Thermo Fisher Scientific). The cells were gently mixed and incubated at room temperature for 15 minutes. For assessment of apoptosis, 400 mL of 1× binding buffer was then added to each tube and the samples were analyzed by flow cytometry using a LSRII instrument (Becton, Dickinson and Company, Franklin Lakes, NJ, USA).
The induction of apoptosis was also monitored by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. The TUNEL assay was performed according to the guidelines recommended by the TUNEL Apoptosis Kit (R&D Systems, Inc.).
The induction of apoptosis was also monitored by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. The TUNEL assay was performed according to the guidelines recommended by the TUNEL Apoptosis Kit (R&D Systems, Inc.).
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