Description
Application
Specificity
Far Red fluorescence detection reagents are often used with Cy®5 filter.
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® technologythat 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 Duolink® PLA Probemaker Guide for Multicolor Detection and the Duolink® PLA Multicolor Detection protocol to use this product.
Detect multiple PPI, PTM and low abundant protein events in a single sample.
Visualize protein complex formation and dissociation
Monitor protein signaling pathways
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 Probemaker Kit - Far Red contains all the necessary reagents to perform the conjugations of Far Red Oligo H and Far Red Oligo I directly to primary antibodies. The resulting PLA probes (Far Red H and Far Red I) must be used together and cannot be paired with any other Duolink® PLA probe.
This product is comprised of the following:
Far Red Oligo H DUO86040H
Far Red Oligo I DUO86040I
Conjugation Buffer DUO82033
Stop Reagent DUO82034
Storage Solution DUO82035
See datasheet for more information.
To perform a Multicolor Duolink® PLA experiment primary antibodies (PLA, IHC, ICC or IF validated) that recognize protein targets of interest are required. Duolink® PLA Multicolor Probemaker-generated probes (Red A/Red B, Green C/Green D, Orange F/Orange G, and/or FarRed H/FarRed I) are used in combination to perform the Multiplex experiment.The reagents in the Duolink® PLA Multicolor Reagent Pack are required to perform the amplification and detection of the resulting PLA probes. Required 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
