Click-iT Plus TUNEL Alexa Fluor 488, 594 and 647 Assay Kits for In Situ Apoptosis Detection enable detection by incorporating smaller highly specific labeling moieties and brighter fluorescent dyes Apoptotic cells in tissue and cultured cell samples. After the labeled moiety is incorporated into the DNA fragment, detection is achieved by catalyzing a "click" reaction under conditions that are mild enough to preserve the fluorescent signal of GFP or RFP.
Additional benefits of the Click-iT Plus TUNEL Assay Kit for In Situ Apoptosis Detection include:
• Optimized to detect apoptotic cells in tissue or cell samples
• Supports multi-channel detection—optimized for use with fluorescent dyes or proteins such as GFP and RFP
• Improved TUNEL detection—enables better label incorporation due to smaller reaction portions
• Bright apoptotic signal—uses Alexa Fluor dye to produce a stable, non-photobleachable fluorescent signal
• Flexibility—the assay can be configured to detect 1–50 samples at a time
Cellular DNA Fragmentation is a hallmark of apoptosis. TUNEL detection is a common method for detecting apoptotic cell DNA fragments in tissue samples. The TUNEL assay first incorporates modified dUTP at the 3'-OH end of the DNA fragment. dUTP modification usually involves the addition of a fluorophore. Due to the different sizes of fluorophores, the incorporation rate of modified dUTP is lower than the expected binding rate, which affects the sensitivity of TUNEL detection. In addition, the multiple fluorophores used in currently available TUNEL assay kits suffer from photobleaching and fluorescence spectral overlap, both of which reduce the sensitivity and multi-channel detection capabilities of the assay.
The Click-iT Plus TUNEL Test Kit was developed to address these issues. The assay uses dUTP modified with an alkyne group, a small bioorthogonal functional group, which allows for easier incorporation of nucleotides. Upon incorporation, a highly specific click reaction is generated between the alkyne group and the Alexa Fluor pyridylmethylazide fluorescent dye, which is a sensitive dye for detecting apoptotic cells or tissue samples. and specific detection. Due to the mild reaction conditions of the Click-iT Plus TUNEL Assay Kit, the assay can also be used with fluorescent proteins or dyes for multi-channel detection.
The Click-iT Plus TUNEL Assay Kit has been validated using several different formalin-fixed paraffin-embedded tissue types. In all cases, the multi-pass detection performance of fluorescent proteins and dyes was retained. Additionally, the ability to stain actin with fluorescently labeled phalloidin is retained.
The Click-iT Plus TUNEL Assay Kit contains all the reagents needed to detect apoptotic cells in tissue or cell samples. The reagents provided in this kit can be used to detect 50 samples and can be configured to detect 1–50 samples at a time.
For Research Use Only. Not for use in diagnostic procedures.