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DUO82040

Sigma-Aldrich

Duolink® In Situ Mounting Medium with DAPI

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Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
NACRES:
NA.32

product line

Duolink®

Quality Level

technique(s)

proximity ligation assay: suitable

fluorescence

λex 360 nm; λem 460 nm (Zeiss Filter set 49)

suitability

suitable for fluorescence

shipped in

wet ice

storage temp.

2-8°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.

Use the Duolink® In Situ Fluorescence Protocol for 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
Duolink® In Situ Mounting Medium with DAPI is ideal for nuclear staining and preserving signals generated with the Duolink® In Situ Detection Reagents for fluorescence microscopy. See datasheet for more details.
Note: Counterstaining with Cy®2 is not recommended.

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

View full Duolink® product list
Duolink® In Situ mounting medium with DAPI has been used to mount coverslips in proximity ligation assay (PLA) for nuclear staining.

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

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

related product

Product No.
Description
Pricing

Pictograms

Corrosion

Signal Word

Warning

Hazard Statements

Precautionary Statements

Hazard Classifications

Aquatic Chronic 3 - Met. Corr. 1

Storage Class Code

8A - Combustible, corrosive hazardous materials

WGK

WGK 2


Certificates of Analysis (COA)

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Detection of Receptor Heteromerization Using In Situ Proximity Ligation Assay
Gomes I, et al.
Current Protocols in Pharmacology / Editorial Board, S.J. Enna (editor-in-chief) ... [Et al.], 2-16 (2016)
S6K2-mediated regulation of TRBP as a determinant of miRNA expression in human primary lymphatic endothelial cells
Warner MJ, et al.
Nucleic Acids Research, 44(20), 9942-9955 (2016)
Intraneuronal Amylin Deposition, Peroxidative Membrane Injury and Increased IL-1β Synthesis in Brains of Alzheimer?s Disease Patients with Type-2 Diabetes and in Diabetic HIP Rats
Verma N, et al.
Journal of Alzheimer'S Disease, 53(1), 259-272 (2016)
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
Ivan Matic et al.
Molecular cell, 39(4), 641-652 (2010-08-28)
Reversible protein modification by small ubiquitin-like modifiers (SUMOs) is critical for eukaryotic life. Mass spectrometry-based proteomics has proven effective at identifying hundreds of potential SUMO target proteins. However, direct identification of SUMO acceptor lysines in complex samples by mass spectrometry

Articles

Proteins are complex biological molecules essential for cellular structure and functions. The majority of proteins commonly interact with various molecules, including other proteins, in order to exert their functions.

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.

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

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

Protocols

Duolink® PLA Multicolor Detection Protocol

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

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

Related Content

Protein and nucleic acid interaction reagents and resources for investing protein-RNA, protein-DNA, and protein-protein interactions and associated 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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