Duolink System
The Duolink® in situ Proximity Ligation Assay (PLA™) from Olink Bioscience enables the in-situ detection and visualisation of individual proteins, protein modifications and protein complexes in tissue and cell samples prepared for microscopy.
In situ PLA Technology
The target is recognised using two antibodies, which are conjugated with oligos which when in close proximity, combine to form a DNA template which can be amplified by rolling circle amplification. The detection kits include either fluorescent- or HRP-tagged oligos which hybridise with the amplified DNA. The signal from each detected pair of primary antibodies is then visualised as individual spots. The number of detection events can be quantified (counted) and assigned to a specific sub-cellular location based on microscopy images.
Applications
1. Detection & Quantification of Protein Interactions
2. Detection & Quantification of Protein Modification
3. Detection & Quantification of a Single Protein Target with High Sensitivity
4. Detection & Quantification of a Single Protein Target with High Specificity
1. Detection & Quantification of Protein Interactions
When detecting two interacting proteins two different primary antibodies are required, one against each interacting protein. The primary antibodies must be raised in two different species. For a complete Duolink® assay, one PLA probe PLUS and one PLA probe MINUS against the different species together with Duolink® Detection Reagents are required.
2. Detection & Quantification of Protein Modification
To be able to detect and quantify a modification, one primary antibody against the protein and one primary antibody against the specific modification site will be required. The two primary antibodies must be raised in two different species. For a complete Duolink assay, one PLA probe PLUS and one PLA probe MINUS against the different species, together with Duolink Detection Reagents are required.
3. Detection & Quantification of a Single Protein Target with High Sensitivity
In order to detect and quantify protein expression with high sensitivity, one primary antibody is required. In this assay, PLA probes PLUS and MINUS will detect the same primary antibody, that is binding to the protein of interest. One PLA probe PLUS and one PLA probe MINUS against the species that the primary antibody is raised in, together with Duolink Detection Reagents are required.
4. Detection & Quantification of a Single Protein Target with High Specificity
This approach is used when there is a need to detect a single protein target with high specificity and sensitivity. For this application two different primary antibodies, raised in two different species against two epitopes on the same protein are required. For a complete Duolink assay, one PLA probe PLUS and one PLA probe MINUS against the different species together with Duolink Detection Reagents are required.
The Duolink system:
Brochures & Manuals
To download the Duolink Brochure (PDF), please click here.
To download the Duolink II Fluorescence User Manual (PDF), please click here.
To download the Duolink II Brightfield User Manual (PDF), please click here.
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