TUNEL Assay Kit (Enhanced TMR Red)

(No reviews yet) Write a Review
SKU:
ES00335
€329

Description

Background

Assay Genie TUNEL Apoptosis Assay Kit is a highly sensitive, rapid and simple method for cell apoptosis detection. After Biotin-dUTP labeling and DAB staining, cell apoptosis can be detected under ordinary optical microscopy.
This kit is developed to detect tissue (Paraffin embedding, freezing and ultrathin section) and cells (Cell smears, cell climbing smears) in situ apoptotic detection.

Detection Principle

When cells undergo apoptosis, endonuclease enzymes are activated to cleave the genomic DNA between nucleosomes. When genomic DNA is cleaved, the exposed 3'-OH can be added with Tetramethylrhodamine-deoxyuridine triphosphate (TMR Red-dUTP) due to the catalysis of Terminal Deoxynucleotidyl Transferase (TdT), apoptosis cells can be detected by fluorescence microscopy. This is the principle of TUNEL (TdTmediated dUTP Nick-End Labeling) for detecting apoptosis.
The Enhanced Fluorescein Mix in the kit contains TMR Red and fluorescence enhancement factor. The fluorescence enhancement factor can bind to TMR Red and enhance the stability amplifies of the signal, which makes the fluorescence brighter and has stronger anti-quenching ability.

Kit Components:

Components 20T 50T 100T Storage

TdT Equilibration Buffer (5x)

700μl

900μL x 2

3.6 mL

-20°C

TdT Enzyme

20 μL

50 μL

100 μL

-20°C

Proteinase K (100 ×)

20 μL

50 μL

100 μL

-20°C

Enhanced Fluorescein Mix

100 μL

250 μL

500 μL

2-8°C

DNase I (2 U/μL)

5 μL

13 μL

25 μL

-20°C

DNase I Buffer (10 x)

100 μL

250 μL

500 μl

-20°C

Reagents needed but not included:

1. Cell Sample
- Fixative Buffer (Polyformaldehyde dissolved in PBS with final concentration of 4%)
- Permeablilization Buffer (0.1% Triton-100)

2. Paraffin Embedding
- Xylene, ethanol, PBS

3. Freezing Section
- Fixative Buffer (Polyformaldehyde dissolved in PBS with final concentration of 4%)

4. Other Reagents
- PBS, ddH2O, DAPI
- Anti-Fluorescence Quenching Agent

Storage:

Store at -20°C. Enhanced Fluorescein Mix should be stored in dark

Cautions:

1. For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.
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. Prepare the TdT Enzyme working solution before use, it can be store on ice for short time, enzyme activity may lose after long-term preservation.
5. If the 20×DAB Concentrate became dark purple, please do not use it.
6. 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.

View AllClose