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iClick™ EU Andy Fluor™ 488 Imaging Kit

Introduction

Our iClick™ EU Andy Fluor™ 488 Imaging Kit provides a simple and robust assay for detection of global RNA transcription temporally and spatially in cells and tissues. The ability to detect newly synthesized RNA or changes in RNA levels resulting from disease, environmental damage or drug treatments is an important aspect of toxicological profiling. In this assay, newly synthesized RNA can be detected in a simple, two-step procedure. In step one, the alkyne-modified nucleoside, 5-ethynyl uridine (EU), is fed to cells or animals and actively incorporated into nascent RNA. The small size of the tag enables efficient incorporation of the modified nucleoside into RNA, but not into DNA. Detection utilizes the“click" reaction between an azide and an alkyne where the modified RNA is detected with a corresponding azide-containing dye. The assay is compatible with antibody-based detection for multiplex imaging.

Features

  • Simple: Labeling is complete in two steps
  • Fast: Results in as little as 90 minutes
  • Multiplex: Compatible with cell cycle dyes and antibody-based stains
  • Quick and Simple Protocol

 

Specifications

Platform: Fluorescence Microscope
Detection Method: Fluorescent
Ex/Em: 495/520 nm

Applications

Detection of RNA transcription in vitro and in vivo.

 

 

 

 

 

 

 

 

 

 

 

 

Figure 1. Detection of newly synthesized RNA in cell. A549 cells were treated with 100 µM EU for 2 hr, then detected with Andy Fluor™ 488 azide (green), cells were counterstained with DAPI (blue).

Figure 2. Detection of newly synthesized RNA in mouse tissue. A mouse was injected intraperitoneally with 50 mg EU per kilogram body weight, then sacrificed at 0, 12, and 24 hr. The mouse intestine tissue was detected with Andy Fluor™ 488 azide (green), and counterstained with DAPI (blue).

To Order

Buy Cat.No. Product Name Ex/Em Unit Price
A009-100T iClick™ EU Andy Fluor™ 488 Imaging Kit 495/520 100 assays 1200
A009-250T iClick™ EU Andy Fluor™ 488 Imaging Kit 495/520 250 assays 2600

Documents

Protocol (PDF): A009
MSDS (PDF):  MSDS-A009
COA (PDF): COA-A009

Reference:

Visualization of actin filaments and monomers in somatic cell nuclei.
Belin BJ, Cimini BA, Blackburn EH, Mullins RD,
Mol Biol Cell (2013) 24:982-994

Bookmarking by specific and nonspecific binding of FoxA1 pioneer factor to mitotic chromosomes.
Caravaca JM, Donahue G, Becker JS, He X, Vinson C, Zaret KS,
Genes Dev (2013) 27:251-260

Visualizing coronavirus RNA synthesis in time by using click chemistry.
Hagemeijer MC, Vonk AM, Monastyrska I, Rottier PJ, de Haan CA,
J Virol (2012) 86:5808-5816