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PLA and Single Cell Proteomics

National facility

The facility employs proximity-based assays for high-specificity protein analyses. Antibodies conjugated to DNA oligonucleotides are used as affinity reagents to allow very sensitive protein detection with a high capacity for multiplexing.

 

SERVICES

  • In situ proximity ligation assays (PLA) in fixed cells or tissue sections to image, with enhanced specificity, the presence of specific proteins, interactions and/or post-translational modifications to e.g. characterise acts of protein signalling in tumor tissues, or to evaluate effects of drug treatment.
  • PLA to detect surface proteins of cells or extracellular vesicles using flow cytometric readout.
  • PLA to increase sensitivity and specificity of Western blot experiments.
  • Other proximity based single- or multiplex assays with dual or triple recognition of the target protein for highly sensitive and specific protein detection in solutions such as plasma and cell/tissue lysate.
  • Conjugation of oligonucleotides to antibodies for custom single-binder or proximity-based protein assays.
  • Access to state-of-the art methods to detect proteins, or proteins combined with mRNA transcripts, in single cells.

 

APPLICATIONS

  • Visualise and quantify proteins, their interactions and post-translational modifications in many types of samples including fixed cells and tissues, cell and tissue lysates, and plasma.
  • Analyse biological pathways and cell signalling.
  • Study extracellular vesicles based on protein content or surface proteins.
  • Investigate heterogeneity in cell populations or tissues at the level of protein (possibly in combination with mRNA).
  • Validate or follow-up findings from transcriptomic studies and investigate RNA-protein expression dynamics.
  • Quantitatively analyse of the effect of drug candidates on protein expression, interactions and modifications.

EQUIPMENT

  • Microscopes:
    • epifluorescence
    • confocal
    • automated scanning
  • C1 Single-Cell Auto Prep system, Fluidigm
  • Realtime qPCR instrument for standard 96- and 384-well plates
  • Access to BioMark HD system, Fluidigm
  • Plate reader for absorbance and fluorescence detection