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Application

SpecificityFluorescence detection reagents are often used with FITC, Cy^®3,Texas Red^® or Cy^®5 filter. The filter chosen is dependent on the color of the Multicolor Probemaker kit selected.Application NotePrimary antibodies 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.Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink^® projectsView full Duolink^® product list

Features and Benefits

Multicolor Duolink^® proximity ligation assay (PLA^®) is an application of the Duolink^® technology that allows for endogenous, simultaneous detection of up to 4 protein interactions, post-translational modifications, or protein expression levels (including low abundant protein) in fixed cells and tissue samples. Follow the Multicolor Duolink PLA protocol to use this product. Visualize and detect multiple proteins events in one experiment. No overexpression or genetic manipulation required High specificity (fewer false positives) Detection of low abundant proteins due to rolling circle amplification Relative quantification possible No special equipment needed Faster and more simple than FRET Increased accuracy compared to co-IP Publication-ready resultsComponentsDuolink^® PLA Multicolor Reagent Pack contains all the necessary reagents to perform the amplification and detection of bound custom PLA probes generated using Duolink^® PLA Multicolor Probemaker Kits. Sufficient components are provided for the indicated number of reactions (30 or 100 RXN), based on 40 mL of the total reaction mixture covering 1 cm². This product is comprised of the following: 5X Multicolor PLA LIgation Buffer DUO86002 Ligase DUO82027 or DUO82025 5X Amplification Buffer DUO82050 Polymerase DUO82028 or DUO82026 5X Multicolor PLA Detection Buffer DUO86001 1X Blocking Buffer DUO82007 1X Probemaker PLA Probe Diluent DUO82036See datasheet for more information.To perform a Multicolor Duolink^® PLA experiment you will need primary antibodies (PLA, IHC, ICC or IF validated) that recognize target epitopes. You will also need at least two pair of the resulting Duolink^® PLA Multicolor Probemaker-generated probes (Red A/Red B, Green C/Green D, Orange F/Orange G, and/or FarRed H/FarRed I).The reagents in the Duolink^® PLA Multicolor Reagent Pack are required to perform the amplification and detection of bound Multicolor PLA probes. Recommended additional reagents include Duolink^® Wash Buffers and Mounting Medium.

Legal Information

Cy is a registered trademark of Cytiva

Duolink is a registered trademark of Sigma-Aldrich Co. LLC

Texas Red is a registered trademark of Life Technologies

PLA is a registered trademark of Sigma-Aldrich Co. LLC