Description
Application
Specificity
Fluorescence 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 Note
Primary 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.
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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 results
Components
Duolink® 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 DUO82036
See 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 Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany
Texas Red is a registered trademark of Life Technologies
