Description_x000D_ Application_x000D_ 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._x000D_ Use the Duolink™ In Situ Fluorescence Protocol for this product. A set of short instructionsis also available._x000D_ Visit our Duolink™ PLA Resource Center for information on how to run a Duolink™ experiment, applications, troubleshooting, and more._x000D_ 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._x000D_ Specificity_x000D_ 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._x000D_ Note: Counterstaining with Cy®2 is not recommended._x000D_ Application Note_x000D_ 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._x000D_ Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink™ projects_x000D_ View full Duolink™ product list_x000D_ Duolink™ In Situ mounting medium with DAPI has been used to mount coverslips in proximity ligation assay (PLA) for nuclear staining._x000D_ Features and Benefits_x000D_ • No overexpression or genetic manipulation required_x000D_ • High specificity (fewer false positives)_x000D_ • Single molecule sensitivity due to rolling circle amplification_x000D_ • Relative quantification possible_x000D_ • No special equipment needed_x000D_ • Quicker and simpler than FRET_x000D_ • Increased accuracy compared to co-IP_x000D_ • Publication-ready results_x000D_ Legal Information_x000D_ Cy is a registered trademark of Cytiva_x000D_ Duolink is a trademark of Sigma-Aldrich Co. LLC_x000D_ PLA is a registered trademark of Sigma-Aldrich Co. LLC