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Quantitative, Fluorescence-Based In-Situ Assessment of Protein Expression

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Tumor Biomarker Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 520))


As companion diagnostics grow in prevalence and importance, the need for accurate assessment of in situ protein concentrations has increased. Traditional immunohistochemistry (IHC), while valuable for assessment of context of expression, is less valuable for quantification. The lack of rigorous quantitative potential of traditional IHC led to our development of an immunofluorescence-based method now commercialized as the AQUA® technology. Immunostaining of tissue samples, image acquisition, and use of AQUA software allow investigators to quickly, efficiently, and accurately measure levels of expression within user-defined subcellular or architectural compartments. IHC analyzed by AQUA shows high reproducibility and demonstrates protein measurement accuracy similar to ELISA assays. The process is largely automated, eliminating potential error, and the resultant scores are exported on a continuous scale. There are now numerous published examples where observations made with this technology are not seen by traditional methods.

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Correspondence to David L. Rimm .

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Moeder, C.B., Giltnane, J.M., Moulis, S., Rimm, D.L. (2009). Quantitative, Fluorescence-Based In-Situ Assessment of Protein Expression. In: Tainsky, M. (eds) Tumor Biomarker Discovery. Methods in Molecular Biology, vol 520. Humana Press, Totowa, NJ.

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-810-2

  • Online ISBN: 978-1-60327-811-9

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