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DUO96000

Sigma-Aldrich

Duolink® PLA Multicolor Reagent Pack

Synonym(s):

in situ Proximity Ligation Assay Kit, Protein Protein Interaction Kit

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About This Item

UNSPSC Code:
41105331
NACRES:
NA.41

product line

Duolink®

technique(s)

immunofluorescence: suitable
multiplexing: suitable
proximity ligation assay: suitable

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

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

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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Chatchai Phoomak et al.
Science advances, 7(3) (2021-02-02)
Asparagine (N)-linked glycosylation is required for endoplasmic reticulum (ER) homeostasis, but how this co- and posttranslational modification is maintained during ER stress is unknown. Here, we introduce a fluorescence-based strategy to detect aberrant N-glycosylation in individual cells and identify a

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