MilliporeSigma
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DUO82049

Sigma-Aldrich

Duolink® In Situ Wash Buffers, Fluorescence

Synonym(s):
Anti-MADA, Anti-LMNL1
NACRES:
NA.32

Quality Level

material

packing (powdered buffer pouches)

product line

Duolink®

technique(s)

proximity ligation assay: suitable

suitability

suitable for fluorescence

storage temp.

20-25°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 fluorescence applications use two wash buffers. Wash Buffer A is used after the PLA Probe incubation step and Wash Buffer B is used after incubation with the amplification reagents. See datasheet for more information.

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®PLA 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.

Linkage
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View full Duolink® product list
Duolink® PLA 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).

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

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

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Articles

Duolink® PLA Fluorescence Protocol

Protocol for immunofluorescent detection of proteins in cells and tissue

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.

Duolink® References

Find Duolink references based on the type of method used, post translational modification detected, and research focus.

Protocols

Duolink® PLA Multicolor Detection Protocol

Duolink® PLA Multicolor Detection Protocol

Duolink® In Situ Short Instructions - Fluorescence

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

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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