Fragel dna fragmentation detection kit

Manufactured by Merck Group

Sourced in United States, Germany

The FragEL DNA Fragmentation Detection Kit is a laboratory equipment product used to detect and analyze DNA fragmentation. It provides a method for the identification and quantification of apoptotic cells, which is a key cellular process. The kit includes the necessary reagents and materials to perform this analysis.

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Lab products found in correlation

Axioskop 2 plus, by Zeiss (4 mentions) Neuronal nuclei (neun), by Merck Group (2 mentions) Fluorescent tdt enzyme, by Merck Group (2 mentions) Ab66155, by Abcam (1 mentions) Ab85554, by Abcam (1 mentions) B5002, by Merck Group (1 mentions) A1r multiphoton microscope, by Nikon (1 mentions) Quant centre software, by Sysmex (1 mentions) Hiseq 2500, by Illumina (1 mentions) Milliplex map kit, by Merck Group (1 mentions) Protein a g magnetic bead, by Thermo Fisher Scientific (1 mentions) Ecl reagent, by Bio-Rad (1 mentions) Lps from escherichia coli, by Merck Group (1 mentions) Egf recombinant human protein, by Thermo Fisher Scientific (1 mentions) Poly l lysine solution, by Merck Group (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Nis elements software, by Nikon (1 mentions)

39 protocols using fragel dna fragmentation detection kit

TUNEL Assay for Apoptosis Detection

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Paraffin sections (n = 6 mice/group) of 5-μm thickness that had been mounted on slides were rehydrated and stained using the Fragment End Labeling (FragEL) DNA Fragmentation Detection Kit with the TUNEL method, following the manufacturer's instructions (Millipore, Billerica, MA). This kit allows the recognition of apoptotic nuclei in paraffin-embedded tissue sections fixed on slides by FragEL of DNA. The terminal deoxynucleotidyl transferase binds to exposed ends of DNA fragments generated in response to apoptotic signals, and catalyzes the template-dependent addition of biotin-labeled and biotin-unlabeled deoxynucleotides. Biotinylated nucleotides are detected using streptavidin-HRP conjugate. Diaminobenzidine reacts with the labeled sample to generate an insoluble colored product at the site of DNA fragmentation. Counter-staining with methyl green aids in the morphologic evaluation and characterization of normal and apoptotic cells. Stained apoptotic epithelial cells (a minimum of 10 microscopic fields per section) were counted manually in a single-blind fashion.

Rao C.V., Janakiram N.B., Madka V., Devarkonda V., Brewer M., Biddick L., Lightfoot S., Steele V.E, & Mohammed A. (2015). Simultaneous targeting of 5-LOX-COX and EGFR blocks progression of pancreatic ductal adenocarcinoma. Oncotarget, 6(32), 33290-33305.

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TUNEL Staining for DNA Fragmentation

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For terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, FragEL™ DNA Fragmentation Detection Kit (Millipore) was used according to manufacturer’s instructions.

Yang S., Gao L., Lu F., Wang B., Gao F., Zhu G., Cai Z., Lai J, & Yang Q. (2015). Transcription factor myocyte enhancer factor 2D regulates interleukin-10 production in microglia to protect neuronal cells from inflammation-induced death. Journal of Neuroinflammation, 12, 33.

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Islet Morphometry and Beta Cell Proliferation

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Islet mass was measured on paraffin sections prepared as described [13 (link)] using an anti-chromogranin A antibody (Ab85554; Abcam, Toronto, ON, Canada) to label endocrine cells. Beta cell proliferation was measured by immunohistochemical staining of pancreas cryosections for Ki67 (Ab66155; Abcam) and insulin as described [13 (link)] or by intravenous injection of BrdU (B5002; Sigma-Aldrich, Oakville, ON, Canada) 50 mg/kg at 12, 36 and 60 h during the infusion followed by immunostaining for BrdU (347580; Becton Dickinson, San Jose, CA, USA) and insulin. Beta cell proliferation was expressed as a percentage of double-positive Ki67⁺ insulin⁺ (or BrdU⁺ insulin⁺) cells to total insulin⁺ cells. Apoptosis was measured using the FragEL DNA Fragmentation Detection Kit (Millipore, Etobicoke, ON, Canada) according to the manufacturer’s instructions (see ESM Methods). The experimenter was blind to group assignments.

Moullé V.S., Vivot K., Tremblay C., Zarrouki B., Ghislain J, & Poitout V. (2017). Glucose and fatty acids synergistically and reversibly promote beta cell proliferation in rats. Diabetologia, 60(5), 879-888.

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Quantitative Analysis of TUNEL Staining

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TUNEL assay was performed by using the FragEL DNA Fragmentation Detection Kit (Millipore #QIA39) following the manufacturer’s instructions. After TUNEL staining, sections were then incubated with DAPI-containing mounting media. The 10× objective lens plus 1.5× optical zoom Z-stack images were obtained using the Nikon A1R⁺ multiphoton microscope. Then, the number of TUNEL⁺ and DAPI⁺ cells within the whole image views was quantified by using the Spots function of Imaris. The density (per 1 mm²) and percentage (%) of TUNEL⁺ cells were calculated.

Zhou B., Chen L., Liao P., Huang L., Chen Z., Liao D., Yang L., Wang J., Yu G., Wang L., Zhang J., Zuo Y., Liu J, & Jiang R. (2019). Astroglial dysfunctions drive aberrant synaptogenesis and social behavioral deficits in mice with neonatal exposure to lengthy general anesthesia. PLoS Biology, 17(8), e3000086.

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Histological Analysis of Fly Brains

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Fly heads without proboscis and antennas were fixed in fresh
Carnoy’s fixative (ethanol: Chloroform: acetic acid at 6:3:1)
overnight at 4°C. Heads were then consecutively washed at RT with 40,
40, 70 and 100% EtOH for 10 min each; following which they were incubated in
methyl benzoate for 30 min at RT and in methyl benzoate: paraffin at 1:1
ratio for 1 h at 65°C. Heads were then infiltrated with paraffin
twice for 1 h at 65°C and embedded in paraffin blocks. The blocks
were sectioned at a thickness of 5 μm, subjected to hematoxylin and
eosin staining, and examined by brightfield microscopy (Axioskop 2 Plus,
Zeiss). For each genotype, vacuoles were counted from one slice around
mid-brain from 20 single flies. For TUNEL assay, 5 mm paraffin sections of
40-day-old fly heads were stained using the FragEL™ DNA fragmentation
detection kit, fluorescent– TdT Enzyme (Millipore, Sigma) according
to the manufacturer’s instructions. Image analysis was performed
using FIJI software (Schindelin et al.,
2012) or Imaris image analysis software (Bitplane).

Maksoud E., Liao E.H, & Haghighi A.P. (2019). A Neuron-Glial Trans-Signaling Cascade Mediates LRRK2-Induced Neurodegeneration. Cell reports, 26(7), 1774-1786.e4.

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