Antibodies for ACTN2, ANK2, ATXN1, COL1A1, COL4A4, COL6A2, CXCL12, SNCA, VDR, WFS1 were purchased from Abcam (Cambridge, MA, USA). The genotyping primers of these genes were listed in Supplementary Table 1. GAPDH was used as interior references for myocardial tissues, which were also purchased from Abcam (Cambridge, MA, USA). The ROS fluorescent probes and JC-1 fluorescent probes were purchased from Beyotime Biotechnology (Shanghai, CHINA). MitoTracker Deep Red was purchased from Invitrogen (Carlsbad, CA, USA). The TUNEL detection kit was purchased from Roche (Indianapolis, IN, USA). All other chemicals were purchased from Sigma unless specifically mentioned otherwise.
Tunel detection kit
Manufactured by Roche
Sourced in United States, Germany, Switzerland, China
The TUNEL detection kit is a laboratory tool used to detect and quantify apoptosis, a form of programmed cell death, in tissue samples. The kit utilizes the TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay, a method that identifies DNA fragmentation, a hallmark of apoptosis. The kit provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Merck Group (5 mentions)
Fluorescence microscope, by Olympus (3 mentions)
Fluorescent microscope, by Olympus (3 mentions)
Hb050 inverted microscope, by Zeiss (3 mentions)
Annexin 5 apc apoptosis detection kit, by Beyotime (2 mentions)
Hb050 inverted microscope system, by Zeiss (2 mentions)
B27 supplement, by Thermo Fisher Scientific (2 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (2 mentions)
Vegf c, by Roche (2 mentions)
N2 supplement, by Thermo Fisher Scientific (2 mentions)
Dmem f 12 glutamax, by Thermo Fisher Scientific (2 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)
Insulin, by Merck Group (2 mentions)
Eclipse te300, by Nikon (2 mentions)
Primescript rt reagent kit, by Takara Bio (2 mentions)
Fluoview 1000, by Olympus (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Jc 1 fluorescent probe, by Beyotime (1 mentions)
Col1a1, by Abcam (1 mentions)
Cxcl12, by Abcam (1 mentions)
108 protocols using tunel detection kit
1
Molecular Profiling of Cardiac Tissues
2
Detecting PC12 Cell Apoptosis via TUNEL Assay
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PC12 cell apoptosis was detected by TUNEL assays with a TUNEL detection kit according to manufacturer’s instruction (Roche, Germany). In brief, the administration and fixation of PC12 cells with CM and 4% paraformaldehyde were carried out. Following the cultivation with proteinase K for 15 min in 37°C, cells were placed in 3% H2O2. After the rinse with PBS for several times, cells were treated by TUNEL detection kit and images were captured by a fluorescence microscopy (Olympus BX53, Japan). Positive cells were counted, and at least 10 randomly chosen fields for each sample were examined per section.
3
TUNEL Assay for Apoptosis in Glioma Cells
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The apoptotic cells were observed by fluorescence stain using a TUNEL detection kit (Roche, Indianapolis, IN, USA) according to the method described elsewhere.20 (link) The extensive DNA fragmentation of the U87 and U251 cells was featured with condensed nuclei. All the slides and tumor sections were observed in six microscopic fields (400×) by two investigators blind to the grouping.
4
TUNEL Assay for Apoptotic Cell Detection
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The detection of apoptotic cells was performed using TUNEL as previously reported (13 (link)). The tissue blocks were fixed in 4% paraformaldehyde and incubated with proteinase K. Fragments of DNA in the tissue sections were analyzed using a TUNEL detection kit (Roche). For each slide, the color images of 10 separate fields were captured randomly and digitized. The cells with clear nuclear labeling were defined as TUNEL-positive cells. The apoptotic index (AI) was calculated as the number of TUNEL-positive cells/total number of myocytes × 100.
5
TUNEL Assay for Apoptosis in ESCC
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A TUNEL assay was used to assess the apoptosis of KYSE-410 and KYSE-1140 cells which were treated with radiation (8 Gy), US, or radiation with USMBs (high or low concentration). The cells were washed with PBS and treated with 4% paraformaldehyde for 15 min and 0.25% Triton‐X 100 for 20 min at 4°C. Subsequently, KYSE-410 and KYSE-1140 cells were subject to a TUNEL detection kit (Roche, Basel, Switzerland) and then were dyed using DAPI. The TUNEL staining of ESCC cells was observed and analyzed using an Eclipse 80i fluorescence microscopy (Nikon, Tokyo, Japan).
6
TRAIL-Induced Apoptosis Quantification
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GC cells were treated with the different concentrations of TRAIL for 24 h. Then a TUNEL Detection Kit (Roche, Basel, Switzerland) was used to determine cell apoptosis, according to the manufacturer’s protocol. Cell confocal images were acquired with a Zeiss LSM700 system (Carl Zeiss, Jena, Germany). Percentages of apoptotic cells are shown from at least 1000 cells, and data were obtained from three independent experiments.
7
Quantifying Apoptotic Cells Using TUNEL Assay
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The apoptotic cells were conducted by using a TUNEL detection kit (Roche Inc, Indianapolis, USA). The sections were incubated at 37°C with TUNEL reaction fluid for 60 minutes add the digoxigenin‐conjugated dUTP to the 3′‐OH ends of fragmented DNA and washed with PBS three times for 45 minutes followed by blocking with 10% goat serum in 0.1 m Tris for 15 minutes. The slides were covered by microscopic glass with mounting medium after three washes again. DNA was fixed by streptavidin‐ HRP peroxidase (1:40 dilution) and dyeing with DAB Chromogen. The apoptotic cells showed cell atrophy with crimp brown nucleus was performed by ZEISS HB050 inverted microscope. The positive cells were monitored and analysed by two observers blinding to the experiment. TUNEL‐positive cells in four aspects from each sample were elected for quantification. The average percentage of these aspects was confirmed as the ultimately data.
8
Quantifying Apoptosis in Rat Hippocampus
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Cell apoptosis in rat brain was detected by TUNEL according to the manufacturer’s protocol (TUNEL Detection Kit, Roche, USA). Briefly, the paraffin sections were heated and dewaxed, rehydrated, and washed 3 times in phosphate-buffered saline (PBS). Proteinase K was used to digest tissues at 37°C for 15 min. The sections were then incubated with 50 μL of TUNEL reaction mixture at 37°C for 1 h. Sections were incubated with Converter-POD solution at 37°C for 30 min. Positive cells (brown staining) and total cells in the hippocampal CA1 region were counted under a light microscope by 2 independent individuals who were blind to the experimental condition [22 (link)]. At least 5 representative CA1 fields were randomly counted per section and 3 sections were counted per brain (n=4/group). Apoptotic index (positive apoptotic cells/total cells ×100%) was used to measure apoptotic severity [23 (link)].
9
Quantifying Apoptotic Podocytes in Glomeruli
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The apoptotic cells of glomerulus were detected by TUNEL detection kit (Roche Diagnostics, Mannheim, Germany) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's instruction. Renal tissue sections (4 μm) were incubated with protease K for 30 min at 37°C and then washed by 3% H2O2 for 10 min at room temperature. The mixture of terminal deoxynucleotidyl transferase (TdT) and biotin-dUTP was added to the renal sections. After that, the sections were incubated with streptavidin-horseradish peroxidase and then were stained by DAB. The apoptotic cells were observed by fluorescent microscope. Apoptotic cells residing on the outer surface of GBM were considered as podocyte. The total number of apoptotic podocytes in each group was counted by examining 10 glomeruli and then they were used to calculate apoptotic index.
10
Cytotoxicity Evaluation of Serum Treatments
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The main reagents include human serum and fetal calf serum (COOK), dimethyl sulfoxide (DMSO), ethylene glycol (EG), saccharose and penicillin-streptomycin, and other reagents (Sigma), as well as the Ki-67 antigen (Abcam), SP kit (Bioss), and TUNEL detection kit (Roche). There were a total of 54 animals, which comprised of 5–6 year-old female nude mice with a body mass of 18–22 g (Beijing Vital River Laboratory Animal Technology Co., Ltd.). The experimental animal quality conformity certificate was SCXK (J) 2014–0010. Upon approval of the Animal Ethics Committee, these mice were randomly divided into three groups: group A, group B, and group C. Each group comprised of 18 mice.
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