DATE: November 17, 2016 TIME: 8:30am PT, 11:30am ET
The complex interactions between transcription, translation, and post-translational modifications are hidden from view in typical endpoint assays that measure either RNA or protein levels. While in situ hybridization (ISH) provides a method for examining the levels of specific RNA transcripts in individual cells, and immunocytochemistry (ICC) utilizes antibodies to visualize the localization of specific proteins (and protein modifications), the ability to simultaneously observe RNA and protein in a single cell has been thwarted by the incompatibility of ICC and ISH protocols. Further complicating this analysis, traditional ISH methods lack sensitivity for detecting low-abundance RNA species and have limited multiplexing capability that further constrain its compatibility with other cell assays.
This webinar will describe a novel technique combining ICC staining with RNA ISH, utilizing branched DNA signal amplification technology, allowing for single copy mRNA sensitivity. We will demonstrate the utility and versatility of this technique using a variety of antibody and probe combinations relevant for cancer research. We will compare expression of protein and mRNA of HER2, estrogen receptor alpha, and progesterone receptor in several breast cancer cell lines known to express differential levels of these targets. We will also compare expression of several microRNAs known to play a role in cancer development in combination with staining for proteins involved in proliferation, anti-apoptosis, and cell structure. Finally, we will show how this novel technique can be used for other research applications for example the detection of Zika virus RNA in infected cells, analysis of circulating tumor cell models and phenotyping of cells within primary hippocampal cultures.