One-step TUNEL In Situ Apoptosis Kit (Green, FITC)

Introduction

Elabscience® One-step TUNEL In Situ Apoptosis Kit applies a highly sensitive, fast and simple method to detect cell apoptosis. This kit is suitable for in situ apoptosis detection of tissue samples (paraffin section and frozen section) and cells (cell slide and cell smear). The results can be directly observed through a fluorescence microscope.

Product Details

Properties

Application	DNA Fragmentation
Detection method	Fluorometric Method
Sample type	Paraffin section, frozen section, cell slide
Assay time	3 hours
Detection instrument	Fluorescence Microscope
Dye Type	FITC
Ex/Em (nm)	490/520
Channel set	FITC
Filter set	FITC
Other reagents required	4% Paraformaldehyde(E-IR-R113), 0.2% Triton X-100(E-IR-R122), PBS(E-IR-R187), ddH2O,Mounting solution with anti-fluorescence quencher(E-IR-R119), Xylene, Ethanol
Storage	-20°C, shading light
Shipping	Ice bag
Expiration date	12 months

Detection Principle

When cells undergo apoptosis, specific DNA endonucleases will be activated, cutting the genomic DNA between the nucleosomes. The DNA of apoptotic cells is cleaved into multimers of 180~200bp fragments, corresponding to the oligonucleosomal size. Therefore, the DNA of apoptotic cells typically migrates as a ladder of 180~200bp on an agarose gel. The exposed 3'-OH of the broken DNA can be catalyzed by Terminal Deoxynucleotidyl Transferase (TdT) with fluorescein labeled dUTP, which can be detected with fluorescence microscope.

Storage

Store at -20°C for 12 months. Labeling Solution and DAPI Reagent (25 μg/mL) should be stored in the dark.

Notes

This kit is for research use only.
Please take safety precautions and follow the procedures of laboratory reagent operation.
The washing operation should be sufficient, otherwise it will affect the enzyme activity (such as DNase I and TdT Enzyme) subsequent experimental operations. After washing the slides with PBS, please carefully blot the liquid around the sample areas with absorbent paper.
Keep the sample moist during the experiment to prevent the failure of the experiment caused by dry slides.
Avoid repeated freezing and thawing of the Labeling Solution and TdT enzyme. Stirring by vortex is not recommended.
The conditions recommended in this manual are universal. Users can optimize the sample processing time, reagent concentration and other conditions according to different sample types and pre-experiment results, and select the most suitable experimental conditions.

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