Duolink^® PLA Multicolor Probemaker Kit - Far Red

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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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.

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