Name: Fluorescent TUNEL assay kit
SKU: BCTUNL
Price: 695.00 USD
Availability: InStock

Fluorescent TUNEL assay kit

$695.00

Item Cat No.: BCTUNL

Application: Labeling apoptotic cells

Reactivity: Tissue and Cell culture

Fluorescent TUNEL assay kit allows in situ detection and quantification of apoptosis at single cell level on cryosection or FFPE section of human and rodent tissues.

1 Kit (50 slides)

A landmark of apoptosis is the activation of nucleases that degrade the nuclear DNA into fragments of approximately 200 base pairs in length. The method of labeling DNA breaks is referred to as Terminal Deoxynucleotide Transferase dUTP Nick End Labeling, or TUNEL.

The Fluorescent TUNEL assay kit detects the DNA fragmentation of apoptotic cells by exploiting the fact that the DNA breaks expose a large number of 3′-hydroxyl ends. These hydroxyl groups can then be fluorescently labeled by terminal deoxynucleotidyl transferase (TdT), which adds deoxyribonucleotides, including FITC-dUTP, in a template-independent fashion.

The kit contains:

250 ml of Universal antigen retrieval buffer (proprietary)
500 μl of TUNEL Enzyme solution
4.5 ml of TUNEL Label Solution (Fluorescein-dUTP, unlabeled dNTP, potassium cacodylate, CoCl₂ and Tris buffered saline)
250 ml of Permeabilization buffer (Tween-20, NaCl and Tris buffered saline)
1 ml of Mounting buffer (Mowiol and Tris buffered saline)

Store at -20^oC

For Research Use Only. Not for use in clinical diagnostics.

Thaw kit content to room temperature.
Deparaffinize FFPE sections with Xylene.
Rehydrate sections with decreasing concentrations of ethyl alcohol (100% to 70%).
Wash slides 2 times with dH₂O.
Dip slides into universal antigen retrieval buffer.
Microwave sections at boiling temperature for 10min.
Wash slides 2 times with dH₂O.
Dip slides into permeabilization buffer for 20min
Wash slides with 1xPBS
Drain Wash solution against a towel and put slides into the wet chamber.
Add 50 μl of TUNEL Enzyme solution to 450 μl of TUNEL Label Solution to obtain 500 μl of TUNEL reaction mixture. Use 100 μl of TUNEL reaction mixture per slide.
Incubate the slides in the wet chamber at 37^oC for 1hr.
Take slides out of the wet chamber, drain off reaction mixture, and wash three times with 1xPBS.
Add 20 μl of Mowiol solution to the section on each slide. Place a cover glass onto the section and apply a small pressure to remove any trapped air bubble. Let Mowiol solution dry at room temperature for 1hr. Store glass slides at 4^oC for less than 1 month.

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Fluorescent TUNEL assay kit

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