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DUO92001 Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Mouse PLUS

Affinity purified Donkey anti-Mouse IgG (H+L)

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Properties

Related Categories Alphabetical Index, Antibodies, Duolink PLA Probes, Duolink PLA Technology, MO-MR,
biological source   donkey (polyclonal)
antibody form   affinity purified immunoglobulin (secondary antibody)
antibody product type   primary antibodies
species reactivity   mouse
application(s)   immunofluorescence: suitable
  proximity ligation assay: suitable
suitability   suitable for brightfield
  suitable for fluorescence
shipped in   wet ice
storage temp.   2-8°C

Description

Components

This product is comprised of the following:
• 5x PLA Probe Anti-Mouse PLUS - Donkey anti-mouse secondary antibody conjugated to oligonucleotide PLUS
• 1x Blocking Solution - Reagent for blocking of the sample
• 1x Antibody Diluent - For dilution of PLA probes and primary antibodies
See datasheet for more information.

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.

This product can be applied to both the Duolink® In Situ Fluorescence Protocol and the Duolink® In Situ Brightfield Protocol depending on the detection reagents used.

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.HRP is also available for brightfield detection.

Specificity
PLA probe anti-Mouse reacts with whole molecule Mouse IgG and the light chains of other Mouse immunoglobulin′s. The PLA probe anti-Mouse may cross-react with Rat antibodies, but has minimal cross reactivity with bovine, chicken, goat, guinea pig, Syrian hamster, horse, human, rabbit, and sheep serum proteins. A MINUS probe of a different species must be used simultaneously with this product. See our Product Selection Guide 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.

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

Legal Information

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

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


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Protocols & Articles

Articles

Antibody Basics

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Protocols

Duolink® PLA Flow Cytometry Protocol

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Duolink® PLA Product Selection Guide

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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

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Peer-Reviewed Papers
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References

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