Proximity Ligation Assays

Proximity ligation assay are used to detect single molecules

Methods to study posttranslational modifications and protein interactions are essential to determine the activity status of signaling cascades in a cell. To be able to examine mixed cell populations and to study cell communication the analyses need to be performed at a single cell level.

We therefore developed a method for detection of PTM and protein interactions with a single molecule resolution, using microscopy for read-out. The method is called proximity ligation assay (PLA) and includes the recognition of a molecule or complex by two antibodies that are linked to short DNA strands. When the antibodies have bound a circular DNA molecule can be formed, which subsequently can be used to produce a detectable signal. The main advantage with this method is that it can be applied on tissues and cells in clinical specimens, which makes it useful for diagnostic applications.

The aim with our research is to identify the signals governing the growth and survival of cancer cells, by developing a panel of assays, to be used as diagnostic and predictive tools. This knowledge will also be used to develop new therapies to selectively target the PTM and protein interactions that are essential for the tumor cells.

You can read more about our methods at: http://www.igp.uu.se/research/molecular_tools/ola_soderberg/?languageId=1

Latest News

Proximity ligation assay is a protein-analyzing method for detection and characterization of proteins with high specificity and sensitivity.

Author(s): Own Document

Author(s): Ola Söderberg, Karl-Johan Leuchowius, Mats Gullberg, et al

A wide range of affinity reagents can be converted to proximity probes by attaching DNA strands. This protocol describes the creation of such proximity probes by covalent coupling of oligoncleotides to antibodies that can be used as either primary or secondary antibodies in assays.

Author(s): Ola Söderberg, Karl-Johan Leuchowius, Mats Gullberg, et al