The apoptosis of endothelial cells in vein graft was observed using a TUNEL Andy Fluor™ 488 Apoptosis Detection Kit (GeneCopoeia, Rockville, MD, USA). Endothelial cells were labelled according to the manufacturer's instructions. Stained sections were observed using fluorescent microscopy. We counted the nuclei of vascular endothelium (luminal border of the blood vessel). The nuclei of apoptotic cells showed green. The apoptosis rate = the number of apoptotic cells (green)/the number of total cells (purple).
Tunel andy fluor 488 apoptosis detection kit
Manufactured by GeneCopoeia
Sourced in United States
The TUNEL Andy Fluor™ 488 Apoptosis Detection Kit is a laboratory product designed to detect and quantify apoptosis, a form of programmed cell death. The kit utilizes the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) method, which labels DNA strand breaks, a hallmark of apoptosis. The kit employs the Andy Fluor™ 488 dye, which emits green fluorescence upon binding to the labeled DNA fragments. This allows for the visualization and quantification of apoptotic cells using fluorescence microscopy or flow cytometry.
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Lab products found in correlation
Dmi4000b, by Leica (1 mentions)
Histology mounting medium, by Merck Group (1 mentions)
Caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions)
2 7 dichlorofluorescin diacetate dcfda, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Hepes, by Merck Group (1 mentions)
Taurine, by Merck Group (1 mentions)
Sodium bicarbonate nahco3, by Merck Group (1 mentions)
Deme medium, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc assay kit, by Cayman Chemical (1 mentions)
Propidium iodide, by Biotium (1 mentions)
Genejet rna purification kit, by Thermo Fisher Scientific (1 mentions)
4 protocols using tunel andy fluor 488 apoptosis detection kit
1
Endothelial Cell Apoptosis in Vein Grafts
2
Histopathological Evaluation of Liver and Pancreas
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Liver sections from the left lobe were formalin‐fixed, paraffin‐embedded and were stained with haematoxylin/eosin, or Sirius red to assess liver fibrosis. Stained sections were evaluated by an expert pathologist, blind to genotype and study period, following the WHO Classification of Tumours of the Digestive System.21 Pancreas section was formalin‐fixed, paraffin‐embedded and stained with haematoxylin/eosin to quantitatively evaluate Langerhans islets. Apoptosis was assessed by terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick‐end labelling (TUNEL) staining using the TUNEL Andy Fluor™ 488 Apoptosis Detection Kit (Genecopoeia, Maryland, USA) following product instructions. Hepatocyte proliferation was assessed by BrdU immunostatining (Life Technologies, California, USA). Nuclei were counterstained with Hoechst 33 342 and the proportion of TUNEL positive cells was quantified using ImageJ software (National Institute of Health) in an automated fashion.22
3
Detection of Apoptosis in Carotid Arteries
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Apoptosis of VSMCs in ligated carotid arteries was detected using a TUNEL Andy Fluor™ 488 Apoptosis Detection Kit (GeneCopoeia A050) according to manufacturer's instructions. Briefly, sections were deparaffinized and rehydrated, permeabilized by Proteinase K solution, incubated with TdT reaction cocktail, and labeled with Andy Fluor™ 488-Streptavidin staining solution. Sections were mounted with histology mounting medium (Sigma) supplemented with 40,6-diamidino-2-phenylin-dole (DAPI; H-1200, VECTASHIELD) for counterstaining DNA. Sections incubated with TdT reaction cocktail without terminal transferase were used as negative controls. Images were taken by a confocal microscope (DMI 4000B; Leica Microsystems, Wet-zlar, German) and quantitated by Image J software as described previously [27] (link).
4
Cytotoxicity and Apoptosis Assays of Chemicals
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DEME medium, DPBS/modifite, and trypsin were purchased from Gibco (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from Hyclone Laboratories, Inc. (Logan, UT, USA). Taurine, HEPES, sodium bicarbonate (NaHCO 3 ), SYBR, BaP, 2,7-dichlorofluorescin diacetate (DCFDA), and penicillin-streptomycin sodium were purchased from Sigma (St. Louis, MO, USA). The CytoScanTM WST-1 cell cytotoxicity assay kit was purchased from G-Biosciences (USA). The GeneJET RNA purification kit, caspase-3/7 green detection reagent, Thermo Scientific maximum first-strand cDNA synthesis kit, and primers were purchased from Thermo Fisher (USA). The TUNEL Andy Fluor™ 488 apoptosis detection kit was purchased from Gene Copoeia™ (USA). A quick apoptosis DNA ladder detection kit was purchased from Biovision Biotechnology (Toronto, Ontario, Canada). Rhodamine 123 and propidium iodide (PI) were purchased from Biotium (USA). The Annexin V-FITC assay kit was purchased from Cayman Chemical (USA). Goat anti-mouse immunoglobulin G (IgG) and goat anti-rabbit IgG antibodies were purchased from Arigo (USA). Other chemicals used in the study were of cell culture grade and were purchased from common sources.
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