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DUO92007

Sigma-Aldrich

Duolink® In Situ Detection Reagents Orange

Synonym(s):

in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent

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About This Item

UNSPSC Code:
12352200
NACRES:
NA.32

product line

Duolink®

technique(s)

proximity ligation assay: suitable

fluorescence

λex 554 nm; λem 576 nm (Cyanine 3; Zeiss Filter set 20)

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

Application

Duolink®proximity ligation assay(PLA®) allows for endogenous detection of protein interactions, post translational modifications, and protein expression levels at the single molecule level in fixed cells and tissue samples.

Follow the Duolink® In Situ Fluorescence Protocolto use this product. A set of short instructionsis also available.

Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.

To perform a complete Duolink® PLA in situ experiment you will need two primary antibodies (PLA, IHC, ICC or IF validated) that recognize two target epitopes. Other necessary reagents include a pair of PLA probes from different species (one PLUS and one MINUS), detection reagents, wash buffers, and mounting medium. Note that the primary antibodies must come from the same species as the Duolink® PLA probes. Analysis is carried out using standard immunofluorescence assay equipment.
Specificity
Orange fluorescence detection reagents are often used with Cyanine 3 filter.

Application Note
Two primary antibodies raised in different species 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.

Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects

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Features and Benefits

  • No overexpression or genetic manipulation required
  • High specificity (fewer false positives)
  • Single molecule sensitivity due to rolling circle amplification
  • Relative quantification possible
  • No special equipment needed
  • Quicker and simpler than FRET
  • Increased accuracy compared to co-IP
  • Publication-ready results

Components

This product is comprised of the following:
  • 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
  • 1x Ligase (1 unit/μL)
  • 1x Polymerase (10 units/μL)
  • 5x Amplification Orange - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase. It also contains oligonucleotide probes labeled with a fluorophore that hybridize to the RCA product.
See datasheet for more information.

Not included in Detection kit:

Primary antibodies, PLA probes, wash buffers, mounting medium

Legal Information

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany

pictograms

Health hazard
GHS08

signalword

Danger

hcodes

H334

Hazard Classifications

Resp. Sens. 1

Storage Class

10 - Combustible liquids

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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Tomokazu Sumida et al.
Nature immunology (2018-10-31)
Foxp3+ regulatory T cells (Treg cells) are the central component of peripheral immune tolerance. Whereas a dysregulated Treg cytokine signature has been observed in autoimmune diseases, the regulatory mechanisms underlying pro- and anti-inflammatory cytokine production are elusive. Here, we identify
M Aubele et al.
British journal of cancer, 103(5), 663-667 (2010-08-12)
Protein tyrosine kinase 6 (PTK6; breast tumour kinase) is overexpressed in up to 86% of the invasive breast cancers, and its association with the oncoprotein human epidermal growth factor receptor 2 (HER2) was shown in vitro by co-precipitation. Furthermore, expression
Jean Philippe Arnault et al.
Clinical cancer research : an official journal of the American Association for Cancer Research, 18(1), 263-272 (2011-11-19)
The emergence of skin tumors in patients treated with sorafenib or with more recent BRAF inhibitors is an intriguing and potentially serious event. We carried out a clinical, pathologic, and molecular study of skin lesions occurring in patients receiving sorafenib.
Kan V Lu et al.
Cancer cell, 22(1), 21-35 (2012-07-14)
Inhibition of VEGF signaling leads to a proinvasive phenotype in mouse models of glioblastoma multiforme (GBM) and in a subset of GBM patients treated with bevacizumab. Here, we demonstrate that vascular endothelial growth factor (VEGF) directly and negatively regulates tumor
Michaela Graf et al.
Human reproduction (Oxford, England), 26(1), 59-66 (2010-11-30)
Dysfunction of cellular processes in the testes can lead to infertility, tumourigenesis or other testicular disorders. 14-3-3 proteins are known to play pivotal roles in cellular communication, signal transduction, intracellular trafficking, cell-cycle control, transcription and cytoskeletal structure and have been
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Articles

How Proximity Ligation Assays (PLA) Work

Learn how Proximity Ligation Assay technology works and how the protein-protein interaction control kit can confirm in situ detection of EGF-induced EGFR-HER2 dimerization.

How to Optimize the Duolink® Proximity Ligation Assay

Things to consider for preparation, setup and execution of the Duolink® assay protocol

Duolink® PLA Troubleshooting Guide

Support information including tips and tricks, frequently asked questions, and basic troubleshooting.

Protocols

Duolink® In Situ Short Instructions - Fluorescence

This page details the Duolink® In Situ Short Protocol for fluorescence detection

Duolink® PLA Fluorescence Protocol

This protocol describes how to perform immunofluorescent detection of proteins in cells and tissue.

Related Content

Duolink® PLA Applications

Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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