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.
This product can be applied to both the Duolink® In Situ Fluorescence Protocol
and the Duolink® In Situ Brightfield Protocol
depending on the detection reagents used.
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.HRP is also available for brightfield detection.Specificity
Duolink® In Situ
Microplate Heat Transfer Block will increase staining efficiency and reproducibility between different plates and reduce edge effects when using Duolink®
reagents in microtiter plates.
Keep the Heat Transfer Block preheated at 37°C and keep it in the 37°C incubator throughout the whole Duolink® In Situ
protocol. Place the microtiter plate in the Heat Transfer Block during incubation steps.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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