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 instructions
is 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
fluorescence applications use two wash buffers. Wash Buffer A is used after the PLA Probe incubation step and Wash Buffer B is used after incubation with the amplification reagents. See datasheet for more information.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®
PLA 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.Linkage
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