Abstract
Digital proximity ligation assay (PLA) detects single protein molecules with a pair of oligonucleotide-conjugated antibodies and digital PCR (dPCR) readout, which allows absolute quantitation of proteins in single cells with high sensitivity. The pipeline also allows simultaneous measurement of protein and mRNA from the same single cell. The sensitivity of the assay has been further improved with implementation of the assay on a microfluidic system, which enables quantitation of rare protein species, with expression level as low as ~3000 protein molecules per cell.
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References
Albayrak C et al (2016) Digital quantification of proteins and mRNA in single mammalian cells. Mol Cell 61(6):914–924
Fredriksson S et al (2002) Protein detection using proximity-dependent DNA ligation assays. Nat Biotechnol 20(5):473
Gullberg M et al (2004) Cytokine detection by antibody-based proximity ligation. Proc Natl Acad Sci U S A 101(22):8420–8424
Darmanis S et al (2016) Simultaneous multiplexed measurement of RNA and proteins in single cells. Cell Rep 14(2):380–389
Lin J et al (2019) Ultra-sensitive digital quantification of proteins and mRNA in single cells. Nat Commun 10(1):1–10
Kellogg RA et al (2014) High-throughput microfluidic single-cell analysis pipeline for studies of signaling dynamics. Nat Protocol 9(7):1713
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Lin, J., Tay, S. (2022). Ultra-Sensitive Quantification of Protein and mRNA in Single Mammalian Cells with Digital PLA. In: Ooi, A.T. (eds) Single-Cell Protein Analysis. Methods in Molecular Biology, vol 2386. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1771-7_11
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DOI: https://doi.org/10.1007/978-1-0716-1771-7_11
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1770-0
Online ISBN: 978-1-0716-1771-7
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