In this pre-print the entire PICO workflow is introduced and the major PICO features, including sensitivity, specificity, and the multiplex capabilities, are described. Among many examples, HER2, a breast cancer oncoprotein, and the phosphorylation status of 4EBP1 is used to showcase the absolute quantitative feature of PICO.
In this paper PICO* was used to confirm the CRISPR/Cas9 mediated knockout of RANKL in mesenchymal stem cells. PICO enabled the distinction between homozygous and heterozygous knock-outs from as little as 1,000 cells.
In this collaborative project PICO* was used to confirm the interaction between bat influenza viruses and the major histocompatibility complex class II (MHC-II) human leukocyte antigen DR isotype (HLA-DR).
* Please note that in both papers PICO was called Emulsion Coupling (EMUC).
In this application note the sensitivity of PICO was compared to western blot. HER2 amounts, a breast cancer biomarker, were compared in the HER2-positive BT474 and the HER2-negative MCF7 cell line. PICO outperformed western blot, regarding sensitivity, by ~ 200-fold.
In this application note we detected and quantified ERBB2 (HER2) and ERBB3 (HER3) in addition to their interaction using PICO technology. Using Absolute Quantification (AQ), we could show that PICO is approximately ~200 fold more sensitive than co-immunoprecipitation (coIP).
Therefore, PICO is a powerful technology to quantify protein interactions and thus to monitor the activity of signaling pathways
In this application note we detected and quantified extracellular vesicles (EVs) from serum or cell supernatant using PICO technology. The data showed that PICO enables the detection of CD63+ and CD9/CD63+ EVs from cell supernatant and serum.
Here we detected and quantified 4EBP1 and its phosphorylated form using PICO technology. Using Absolute Quantification (AQ), we could show that upon treatment of BT474 or MCF7 cells with the kinase inhibitor Dactolisib, the phosphorylation of 4EBP1 is decreased ~200 fold.
Therefore, PICO is a powerful technology to quantify post-translational modifications and to monitor the activity of signaling pathways.
In this application note we compared the two PICO quantification methods: the Absolute Quantification (AQ) and the Relative Quantification (RQ). Using HER2 as the model protein we showed that BT474 cells have approximately 120-fold more HER2 than MCF7 cells.