Detect, Localize, and Quantify RNA and lncRNA at the Subcellular Level
Stellaris™ RNA FISH technology is a new Biosearch Technologies product for in situ
RNA detection which enables simultaneous detection, localization, and quantification of individual RNA molecules at the subcellular level. This technique provides a high degree of specificity and sensitivity with a simple protocol and low cost per hybridization.
Stellaris RNA FISH utilizes a set of simple fluorescent probes which bind along the length of target RNA. The combined fluorescence of multiple fluorophores in close proximity is visible and has a distinct spot above background. False negatives are minimized, since only a fraction of the probes are required to bind to the target to produce a positive signal. Likewise, false positives are minimized as off target hybridization contributes only weak and diffuse fluorescence, which is not distinguished above background.
The Stellaris FISH protocol is comparatively simple, and consists of only four steps: sample preparation, hybridization, a wash step, and imaging. No exotic reagents are required, and the entire process can be completed in less than a day. The technique can be used in adherent cells, as well as frozen or formalin fixed paraffin embedded tissues, and can be combined with immunofluorescence antibody assays or run as a multiplexed assay with up to three simultaneous RNA targets.
Stellaris FISH is an effective tool for several key applications. For example, Stellaris probes can be used to reveal up- and down- regulation of gene expression in drug screening studies. Another example is the characterization of tumors by correlating mRNA levels of known oncogenes. Stellaris FISH can be used alongside existing technologies such as qPCR, DNA FISH, IHC (immunohistochemistry), and western blotting to provide complementary information while preserving cell morphology. Scientists can also finally address the stochastic nature of gene expression and visualize RNA through direct detection without isolation, purification, and amplification.
Biosearch Technologies currently offers Stellaris FISH probes manufactured on a custom basis, including software for optimum probe design. In addition, pre-designed, catalogued probe sets are available.
Catalogued products include: SMF-3001-1, SMF-2002-1, SMF-2003-1, SMF-2006-1, SMF-2007-1, SMF-2010-1, SMF-2013-1, SMF-2014-1, SMF-2015-1, SMF-2016-1, SMF-2017-1, SMF-2018-1, SMF-2026-1SMF