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.
Mail address
Uppsala University
PLA and Single Cell Proteomics
IGP / BMC
Box 815
751 08 Uppsala
Visitor address / Deliveries
Uppsala University
PLA and Single Cell Proteomics
BMC, Husargatan 3
751 22 Uppsala