Click-iT™ RNA Alexa Fluor™ 488 Imaging Kit detects temporal and spatial changes in total RNA transcription in cells and tissues (PNAS (2008) 105:15779-84).
The ability to detect newly synthesized RNA or changes in RNA levels caused by disease, environmental insult, or drug treatment is an important aspect of toxicological expression profiling. Utilizing an alkyne-modified nucleoside, 5-ethynyluridine (EU), and powerful click chemistry technology, newly synthesized RNA can be detected in a simple two-step procedure without the use of radioactive labels or antibodies. In the first step, the cell or animal is fed with an alkyne-containing nucleoside and actively incorporated into the nascent RNA. The small size of this tag allows the modified nucleosides to be efficiently incorporated into RNA rather than DNA. The detection utilizes a chemoselective ligation or "click" reaction between azide and alkyne, using the corresponding azide dye to detect modified RNA. With its tiny "footprint," Click-iT™ detection molecules can easily penetrate complex samples and have the potential to be used in multi-pathway analyses, including with other probes, including antibodies for the detection of RNA-interacting proteins, to provide deeper biological insights.