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Ultra-Sensitive Quantification of Protein and mRNA in Single Mammalian Cells with Digital PLA

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Part of the Methods in Molecular Biology book series (MIMB,volume 2386)

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.

Key words

  • Single-cell protein quantitation
  • Proximity ligation assay
  • Microfluidics

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  • DOI: 10.1007/978-1-0716-1771-7_11
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Correspondence to Jing Lin .

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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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