RNAscope® assay is an innovative and proprietary RNA in situ hybridization (ISH) assay based on ACD's patented technology with signal amplification and simultaneous background noise suppression. Unique to this technology, RNAscope® assay delivers quantitative, sensitive and specific molecular detection of RNA species on a cell-by-cell basis with morphological context in a single assay. This enables researchers to visualize which genes are expressed, localize where they are expressed, and quantify the level of expression, thus making it ideal tool to complement technologies like IHC, qPCR or NGS. Our recent development, the BaseScope™ assay, now enables detection of any short RNA targets and exon junctions.
This webinar is targeted towards new and intermediate RNA ISH or IHC users. It is intended to be an introduction and does not require previous training.
This webinar will include the following topics:
De-paraffinization is performed to ensure complete removal of the paraffin from FFPE samples to allow for the probes to penetrate the target RNA after adequate pretreatment.
RNAscope H2O2 step is performed during the RNAscope assay to block endogenous peroxidase enzyme activity to prevent hazy background after detection.Note: Pretreatment 1 refers to RNAscope H2O2 reagent and is the first pretreatment perfomed on your samples for Chromogenic assays.
Target Retrieval step is a heat induced epitope retrieval method that is necessary to reverse the cross-linking caused by the formalin fixation step. Note: Pretreatment 2 refers to Target Retrieval reagent.For an alternative steamer protocol refer to the appendix of User Manual Part 2 Brown and Red.
Protease Plus is a broad spectrum protease that is intended to permeabilize the samples adequately to allow the probes to reach the target mRNA.Note : Pretreament 3 refers to Protease Plus reagent.
ACD provides properietary double "ZZ" oligo probes designed to hybridize to your specific RNA target
RNAscope detection reagents amplify hybridization signals via sequential hybridization of amplifiers.
Chromogenic detection is based on the enzyme substrate reaction which leads to the formation of an insoluble precipitate visualized in the form of punctate dots for the RNAscope assay.
Hematoxylin staining is a counterstain used to provide a contrast to better visualize the signal and to observe the morphology of the sample and identify the localization of the signal.
The tissue dehydration step after the counterstaining step results in removal of excess moisture which provides better tissue morphology and preservation of the signal.
The final step in RNAscope after staining requires the use of mounting media to adhere a coverslip to tissue section or cell smear. This helps protect the sample and the staining from physical damage and helps improve the clarity and contrast of an image during microscopy.
Please contact us directly , by phone or email, for any technical assistance with RNAscope® assay or relevant products.
Complete one of the two forms below and we will get back to you.
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