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DUO92002 Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Rabbit PLUS

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

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Properties

Related Categories Alphabetical Index, Antibodies, Duolink PLA Probes, Duolink PLA Technology, Molecular Biology,
biological source   donkey (polyclonal)
antibody form   affinity purified immunoglobulin (secondary antibody)
antibody product type   primary antibodies
species reactivity   rabbit
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-Rabbit PLUS - Donkey anti-rabbit 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-Rabbit reacts with whole molecule Rabbit IgG and the light chains of other Rabbit immunoglobulin′s. The PLA probe anti-Rabbit 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

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products.Once you log onto our website, you will find the price and availability displayed on the product detail page.You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 orview local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
What epitope on the antibody is recognized by the Duolink PLA Probes?
The Duolink PLA Probes offered are donkey polyclonal antibodies directed to the whole molecule (heavy and light chains). They are affinity purified. The same polyclonal antibody is conjugated to the Plus and Minus Oligos. The Plus and Minus probes can be used together (to detect a single antibody) since they can recognize multiple sites.The anti-mouse IgG PLA labeled antibodies may cross-react with rat but this is not consistent. Do not use this product with rat antibodies. It is best to label anti-rat secondary antibodies with the Duolink Probemaker kit.The anti-goat IgG PLA labeled antibodies have cross-reactivity with sheep IgG and can be recommended for this application.
Has Duolink been used for detection of changes in protein expression?
The following reference cites the use of Duolink to detect changes in protein expression in Human.Raizer et al., Cancer 2010;116:5297305
How many reactions can be performed with the Duolink® In Situ Starter Kit?
All of the starter kits for the Duolink product line will perform 30 reactions using 40 ul per reaction. DUO92101 - Duolink® In Situ Red Starter Kit Mouse/Rabbit DUO92102 - Duolink® In Situ Orange Starter Kit Mouse/Rabbit DUO92103 - Duolink® In Situ Red Starter Kit Mouse/Goat DUO92104 - Duolink® In Situ Orange Starter Kit Mouse/Goat DUO92105 - Duolink® In Situ Red Starter Kit Goat/Rabbit DUO92106 - Duolink® In Situ Orange Starter Kit Goat/Rabbit
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Custom Duolink Services
Protocols & Articles

Articles

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Duolink® References

Duolink proximity ligation assay (PLA) has been used in countless studies in a variety of fields. Browse the following tables to view highlights from the many applications of this technology.
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Protocols

Duolink® PLA Flow Cytometry Protocol

This protocol describes the use of Duolink® PLA reagents for the detection of individual proteins, protein modifications, and protein-protein interactions within cell populations by flow cytometry.
Keywords: Amplification, Centrifugation, Flow cytometry, Microscopy, Peptide synthesis

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

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Peer-Reviewed Papers
15

References

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