iClick™ TUNEL Andy Fluor™ 555 Imaging Assay

Introduction

Internucleosomal cleavage of DNA is a hallmark of apoptosis. DNA cleavage in apoptotic cells can be detected in situ in fixed cells or tissue sections using the terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) method. TUNEL is highly selective for the detection of apoptotic cells but not necrotic cells or cells with DNA strand breaks resulting from irradiation or drug treatment.

In the TUNEL assay, TdT enzyme catalyzes the addition of modified dUTP to the 3’-OH ends of fragmented DNA. The modifications are fluorophores or haptens, including biotin or bromine which can be detected directly in the case of a fluorescently-modified nucleotide, or indirectly with streptavidin or antibodies, if biotin-dUTP or BrdUTP are used, respectively.

The new iClick™ TUNEL Andy Fluor™ 555 Imaging Assay utilize a dUTP modified with an alkyne, a small, bio-orthogonal functional group that enables the nucleotide to be more readily incorporated by TdT than other modified nucleotides such as biotin-dUTP and fluorescein-dUTP. Detection is based on a click reaction, a copper (I) catalyzed reaction between an azide and alkyne (Figure 1). The small size of the Andy Fluor™ azide (MW ∼1,000) compared to that of an antibody (MW ∼150,000) enables effortless penetration of complex samples with only mild fixation and permeabilization required. As a result, when compared to assays using other modified nucleotides, the iClick™ TUNEL Imaging Assay is fast (complete within 2 hours) and is able to detect a higher percentage of apoptotic cells under identical conditions.

Features

Broad application: Suitable for FFPE tissue, frozen tissue and cells.
High sensitivity: Can detect a higher percentage of apoptotic cells.
Bright apoptotic signal: Uses Andy Fluor™ dyes, resulting in a stable, non-photobleaching fluorescent signal.

Figure 1. Detection of apoptosis with the iClick™ TUNEL Imaging Assay.

Specifications

Platform:	Fluorescence Microscopy
Detection Method:	Fluorescent
Ex/Em:	555/575 nm
Product Size:	50 assays
Storage Conditions:	-20 ℃, protect from light
Shipping Condition:	Dry ice

Figure 2. Detection of apoptotic cells in mouse tissue using iClick TUNEL Andy Fluor™ 555 Imaging Assay.

To order

Documents

Protocol (PDF):	A061
MSDS (PDF):

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Wu, D., Zhang, Y., Zhao, C. et al.
J Transl Med 20, 583 (2022). https://doi.org/10.1186/s12967-022-03779-9

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Buy	Cat.No.	Product Name	Ex/Em	Unit	Price
	A061	iClick™ TUNEL Andy Fluor™ 555 Imaging Assay	555/575	50 assay	$420

iClick™ TUNEL Andy Fluor™ 555 Imaging Assay

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