Duolink™ PLA Multicolor Probemaker Kit - Orange Proximity Ligation Assay Multiplexing
Specificity
Orange fluorescence detection reagents are often used with Cy®3 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™ 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 Duolink™ PLA Probemaker Guide for Multicolor Detection and the Duolink™ PLA Multicolor Detection 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
Probemaker Kit - Orange contains all the necessary reagents to perform the conjugations of Orange Oligo F and Orange Oligo G directly to primary antibodies. The resulting PLA probes (Orange F and Orange G) must be used together and cannot be paired
with any other Duolink™ PLA probe.
This product is comprised of the following:
• Orange Oligo F DUO86030F
• Orange Oligo G DUO86030G
• 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 trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC
