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TUNEL Assay Kit (Enhance FITC)[334]

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Price:   $300
Cat Number:  E-CK-A334
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Introduction

Elabscience® TUNEL Apoptosis Assay Kit is a highly sensitive, rapid and simple method for cell apoptosis detection. The result can be detected by fluorescence 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 a fluorescein dUTP (FITC-12-dUTP) due to the catalysis of Terminal Deoxynucleotidyl Transferase (TdT), apoptosis cells can be detected by fluorescence microscopy. This is the principle of TUNEL (TdT-mediated dUTP Nick-End Labeling) for detecting apoptosis.

The Enhanced Fluorescein Mix [E-CK-A334D] in the kit contains FITC-12-dUTP and fluorescence enhancement factor. The fluorescence enhancement factor can bind to FITC and enhance the stability amplifies of the signal, which makes the fluorescence brighter and has stronger anti-quenching ability.


Components


Cat

Products

20 Tests

50 Tests

100 Tests

Storage

 E-CK-A334A

TdT Equilibration Buffer (5 ×)

700 μL

900 μL × 2

3.6 mL

-20°C

 E-CK-A334B

TdT Enzyme

20 μL

50 μL

100 μL

-20°C

 E-CK-A334C

Proteinase K (100 ×)

20 μL

50 μL

100 μL

-20°C

 E-CK-A334D

Enhanced Fluorescein Mix

100 μL

250 μL

500 μL

-20°C

 E-CK-A334E

DNase I(2 U/μL)

5 μL

13 μL

25 μL

-20°C

 E-CK-A334F

DNase I Buffer(10 ×)

100 μL

250 μL

500 μL

-20°C

                                  Manual

 One Copy




Reagent not included

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 (0.1% Triton-100, It is recommended to use Elabscience® E-IR-R122).


2.  Paraffin embedding

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


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, ddH2O, DAPI (It is recommended to use ELabscience® E-IR-R103), Anti Fluorescence Quenching Agent (It is recommended to use Elabscience® E-IR-R119).


Instructions

Proteinase K (100 ×)[E-CK-A334C] is concentrated, diluted with PBS to 1 × working solution before use.

For example: take 1 μL Proteinase K (100 ×)[E-CK-A334C], add to 99 μL PBS, the mixture is 1×Proteinase K working solution.


Storage

Store at -20°C. Enhanced Fluorescein Mix [E-CK-A334D] 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.  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.

If you have any question about the product, please feel free to contact us or leave a message