PLA and Single Cell Proteomics

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

Using the facility

Please contact us via email for more details regarding the  services offered by the PLA and Single Cell Proteomics facility.