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DUO80102 Sigma

Duolink® In Situ Brightfield Mounting Medium

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Properties

application(s)   proximity ligation assay: suitable
storage temp.   20-25°C

Description

Application

Experiments conducted using Duolink in situ reagents can detect and visualize protein interactions, protein expression levels and post translational modifications at the single molecule level in fixed cells and tissue samples.

To perform a complete Duolink in situ experiment you will need two primary antibodies (IHC or IF validated) that recognize two target epitopes. Additional reagents required include a pair of PLA probes, one PLUS and one MINUS , your choice of Detection Reagents. Optional reagents include Wash Buffers and Mounting Medium.

Analysis is carried out using standard immunofluorescence assay equipment. HRP/Novared is also available for bright field detection. Quick and reliable quantification can be performed using the Duolink Image Tool.

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) or immunohistochemistry (IHC) assay to determine the optimal fixation, blocking, and titer conditions.
Duolink in situ reagents are suitable for use on fixed cells, cyt-spin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). No minimum number of cells is required.

Find answers to commonly asked question on our Duolink FAQ page

Legal Information

Duolink is a registered trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

Symbol 
Signal word 
Danger
RIDADR 
UN1307 - class 3 - PG 3 - Xylenes
WGK Germany 
3
Flash Point(F) 
73.4 °F
Flash Point(C) 
23 °C
Protocols & Articles

Articles

Detect, Visualize and Quantify Single Post-Translational Modifications

Duolink® In Situ products enable detection, visualization, and quantification of protein events in tissue and cell samples prepared for microscopy. The in situ PLA® technology, on which these product...
Caleb Hopkins, Product Manager, Sigma® Life Science
Biofiles Vol. 8, No. 11
Keywords: Acetylations, Amplification, Apoptosis, Cancer, Cellular processes, Clinical, Diagnostic, Diseases, Enzyme activity, Gene expression, Glycosylations, Immunohistochemistry, Immunoprecipitation, Microscopy, Nucleic acid hybridization, Peptide synthesis, Phosphorylations, Post translational modifications, Transduction, Ubiquitinations, Western blot

Duolink® PLA Applications

With Duolink® PLA, you can detect, quantify, and visualize protein-protein interactions, post-translational modifications, and low expression protein detection with an easy-to-execute immunodetection...
Keywords: Acylations, Amplification, Biological processes, Buffers, DNA replication, Electrophoresis, Flow cytometry, Gel electrophoresis, Gene expression, Glycosylations, High performance liquid chromatography, Immunohistochemistry, Mass spectrometry, Metabolism, Microscopy, Phosphorylations, Post translational modifications, Sulfations, Transcription, Transduction, Ubiquitinations

Protocols

Duolink® In Situ Short Instructions - Fluorescence

Note: Use open droplet reactions without a cover slip and perform all incubations in a humidity chamber. Use volumes corresponding to your reaction area, see the Reaction Volume Guide at sigma.com/du...
Keywords: Amplification, Confocal microscopy, Peptide synthesis, Phase transitions

Duolink® PLA Product Selection Guide

Getting started with Duolink® PLA is simple. The protocols are very similar to that of IF, IHC, and flow cytometry, however Duolink® PLA offers distinct advantages over these and other traditional me...
Keywords: Amplification, Bacterial conjugations, Buffers, Flow cytometry, Fluorescent microscopy, Gene expression, Immunohistochemistry, Microscopy, Peptide synthesis

Duolink® PLA Troubleshooting Guide

The Duolink® PLA technology for protein detection offers a simple and straightforward protocol that has a set of defined steps. This section will delineate important reminders that will ensure your D...
Keywords: Addition reactions, Amplification, Bacterial conjugations, Buffers, Dehydration reaction, Filtration, Flow cytometry, Immunofluorescence, Immunohistochemistry, Peptide synthesis, Purification, Titrations

How to Optimize the Duolink® Proximity Ligation Assay

To successfully execute a Duolink® PLA experiment, there are some necessary considerations to properly prepare for, setup, and execute the assay protocol.
Keywords: Bacterial conjugations, Buffers, Detergents, Flow cytometry, Fluorescent microscopy, Gene expression, Growth factors, Immunofluorescence, Immunohistochemistry, In Situ hybridization, Microscopy, Nucleic acid hybridization, Phosphorylations, Post translational modifications, Sample preparations

Related Content

Duolink® PLA Custom Services

Duolink® Proximity Ligation Assay (PLA) is a powerful tool that allows in situ detection of endogenous proteins, protein interactions, and protein modifications with high specificity and sensitivity....
Keywords: Flow cytometry, Genetic, Peptide synthesis

Duolink® PLA Resource Center

Understanding protein function in the cell can create solutions to improve human health. For that reason we created the Duolink® PLA Resource Center, to provide insightful educational resources so yo...
Keywords: Flow cytometry, Peptide synthesis

Peer-Reviewed Papers
15

References

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