Duolink^® proximity ligation assay(PLA^®) allows for endogenous detection of protein interactions, post translational modifications, and protein expression levels at the single molecule level in fixed cells and tissue samples.

Use the Duolink^® In Situ Fluorescence Protocol for this product. A set of short instructionsis also available.

Visit our Duolink^® PLA Resource Center for information on how to run a Duolink^® experiment, applications, troubleshooting, and more.

To perform a complete Duolink^® PLA in situ experiment you will need two primary antibodies (PLA, IHC, ICC or IF validated) that recognize two target epitopes. Other necessary reagents include a pair of PLA probes from different species (one PLUS and one MINUS), detection reagents, wash buffers, and mounting medium. Note that the primary antibodies must come from the same species as the Duolink^® PLA probes. Analysis is carried out using standard immunofluorescence assay equipment.

Specificity
Duolink^® In Situ Mounting Medium with DAPI is ideal for nuclear staining and preserving signals generated with the Duolink^® In Situ Detection Reagents for fluorescence microscopy. See datasheet for more details.
Note: Counterstaining with Cy^®2 is not recommended.

Application Note
Two primary antibodies raised in different species are needed. Test your primary antibodies (IgG-class, mono- or polyclonal) in a standard immunofluorescence (IF), immunohistochemistry (IHC) or immunocytochemistry (ICC) assay to determine the optimal fixation, blocking, and titer conditions. Duolink^® in situ reagents are suitable for use on fixed cells, cytospin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). No minimum number of cells is required.

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