TUNEL Assay Kit (Enhance TMR Red)[335]
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| Price: | $300 |
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 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 (TdT-mediated dUTP Nick-End Labeling) for detecting apoptosis.
The Enhanced Fluorescein Mix [E-CK-A335D] 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.
Components
|
Cat |
Products |
20 Tests |
50 Tests |
100 Tests |
Storage |
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E-CK-A335A |
TdT Equilibration Buffer (5 ×) |
700 μL |
900 μL × 2 |
3.6 mL |
-20°C |
|
E-CK-A335B |
TdT Enzyme |
20 μL |
50 μL |
100 μL |
-20°C |
|
E-CK-A335C |
Proteinase K (100 ×) |
20 μL |
50 μL |
100 μL |
-20°C |
|
E-CK-A335D |
Enhanced Fluorescein Mix |
100 μL |
250 μL |
500 μL |
-20°C |
|
E-CK-A335E |
DNase I (2 U/μL) |
5 μL |
13 μL |
25 μL |
-20°C |
|
E-CK-A335F |
DNase I Buffer (10 ×) |
100 μL |
250 μL |
500 μL |
-20°C |
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Manual |
One Copy |
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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-A335C] is concentrated, diluted with PBS to 1 × working solution before use.
For example: take 1 μL Proteinase K (100 ×)[E-CK-A335C], add to 99 μL PBS, the mixture is 1×Proteinase K working solution.
Storage
Store at -20°C. Enhanced Fluorescein Mix [E-CK-A335D] 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.
