Detection principle

When cells undergo apoptosis, specific DNA endonucleases will be activated, cutting the genomic DNA between the nucleosomes. After extraction and loading for electrophoresis, a DNA ladder of 180-200bp can be found in such DNA samples, which is a specific event during cell apoptosis. The exposed 3'-OH of the broken DNA can be catalyzed by Terminal Deoxynucleotidyl Transferase (TdT) with labeled dUTP, which can be detected with microscopy or flow cytometry.

Elabscience® One-step TUNEL Assay Kit is a highly sensitive, fast and simple method for cell apoptosis detection. The test results can be directly observed through a fluorescence microscope. This kit is suitable for in situ apoptosis detection of tissue samples (Paraffin embedding, freezing and ultrathin section) and cells (Cell smears, cell climbing smears) in situ apoptotic detection.

Components

Cat.Products20 Assays50 Assays100 Assays

E-CK-A32ATdT Equilibration Buffer4 mL9 mL9 mL × 2

E-CK-A32BTdT Enzyme100 μL250 μL250 μL × 2

E-CK-A32CProteinase K (100 ×)20 μL50 μL100 μL

E-CK-A320DLabeling Solution100 μL × 2100 μL × 5100 μL × 10

E-CK-A32EDNase I (20 U/μL)5 μL13 μL25 μL

E-CK-A32FDNase I Buffer (10 ×)300 μL700 μL1500 μL

E-CK-A32GDAPI (25 ×) 100μL250μL500μL

Related Reagents

1.Cell sample

Fixative Buffer (Polyformaldehyde dissolved in PBS with final concentration of 4%) (It is recommended to use Elabscience®E-IR-R113).

Permeablilization Buffer (Triton-100 dissolved in PBS with final concentration of 0.2%) (It is recommended to use Elabscience® E-IR-R122).

2.Paraffin embedding

Xylene, ethanol.

3.Freezing section

Fixative Buffer (Polyformaldehyde dissolved in PBS with final concentration of 4%) (It is recommended to use Elabscience® E-IR-R113).

4.Other reagents

PBS (It is recommended to use Elabscience® E-BC-R187).

Anti-Fluorescence Quenching Agent (It is recommended to use Elabscience® E-IR-R119).

Storage

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

Cautions

1.Avoid repeated freezing and thawing of the Label Solution and TdT enzyme, Vortex and mix well before use.

2.After washing the slides with PBS, please wipe dry the liquid around the tissue on the slides with absorbent paper.

3.Keep the sample moist during the experiment to prevent the failure of the experiment caused by dry slices.

4.This kit is for research use only. For your safety and health, please wear lab clothes and gloves. Instructions should be followed strictly, changes of operation may result in unreliable results.