Invitrogen™ Click-iT™ TUNEL Alexa Fluor Imaging Assays for Microscopy & HCS

Description

• Screen apoptotic cells easily and efficiently in just two hours with the Click-iT TUNEL Alexa Fluor 488, 594, and 647 imaging assays
• Intended for use with microscopy and high-content screening (HCS), these TUNEL assay kits use a copper-catalyzed reaction to detect incorporated alkyne-modified dUTPs on terminal DNA ends of fragmented DNA
• Compared with assays using other modified nucleotides, these apoptosis detection kits are fast (complete within two hours) and can detect a higher percentage of apoptotic cells under identical conditions
• Click-iT TUNEL assays also allow multiplexing with surface and intracellular biomarker detection methods, including those that measure fluorescent signal from GFP or RFP
• Apoptosis is marked by defined cellular processes, including cell rounding, blebbing, and DNA fragmentation
• The TUNEL assay is the most widely used analytical method to detect fragmented DNA in apoptotic cells in tissue samples, beginning with the incorporation of modified dUTPs at the 3’-OH ends of fragmented DNA
• The dUTPs often include a fluorophore molecule
• Due to the size of the fluorophore, modified dUTPs can display lower than expected incorporation rates, negatively affecting the sensitivity of the TUNEL assay
• Quite often, fluorophores used in currently available TUNEL assay kits exhibit photobleaching and fluorescent spectral overlap issues, both of which reduce the sensitivity of and ability to multiplex these assays
• The Click-iT Plus TUNEL assay was developed to address these issues and is based on a copper (I) catalyzed (i.e., click) reaction between an azide and alkyne
• The small size of the Alexa Fluor azide, which has a molecular weight of about 1 kDa, enables easier incorporation of the Alexa Fluor 488, 594, or 647 dye into complex samples, as compared to larger (MW ˜100,000 Da) antibodies
• This permits mild fixation or permeabilization and faster assay turnaround time (about two hours) and enables the detection of a higher percentage of apoptotic cells under identical conditions
• Because of its gentle reaction conditions, the Click-iT Plus TUNEL assay can also be multiplexed with fluorescent proteins or dyes
• The Click-iT TUNEL Alexa Fluor imaging assays contain all components needed to accurately and reliably detect apoptosis in adherent cells grown on coverslips or 96-well microplates, and they also include DNase I to generate strand breaks for the positive control.