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DUO96040

Sigma-Aldrich

Duolink® PLA Multicolor Probemaker Kit - Far Red

Proximity Ligation Assay Multiplexing: protein-protein interaction, post-translational modification and low abundant protein events

Synonym(s):

in situ Proximity Ligation Assay, Flowcytometry-PLA, Protein Protein Interaction Kit

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About This Item

UNSPSC Code:
41105331
NACRES:
NA.32

technique(s)

immunofluorescence: suitable
multiplexing: suitable
proximity ligation assay: suitable

fluorescence

λex 646 nm; λem 664 nm (Far Red)
λex 646 nm; λem 664 nm

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

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

Pictograms

Exclamation markEnvironment

Signal Word

Warning

Hazard Statements

Hazard Classifications

Aquatic Chronic 2 - Skin Sens. 1

Storage Class Code

10 - Combustible liquids

WGK

WGK 3


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Articles

Duolink® PLA Multicolor technology and the quantification of protein-protein interactions in fixed tissues and cells.

Protocols

Duolink® PLA Multicolor Detection Protocol

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