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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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NACRES:
NA.32

biological source

donkey (polyclonal)

Quality Level

200

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody product type

primary antibodies

product line

Duolink®

species reactivity

rabbit

technique(s)

immunofluorescence: suitable
proximity ligation assay: suitable

suitability

suitable for brightfield
suitable for fluorescence

shipped in

wet ice

storage temp.

2-8°C

Related Categories

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

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.

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

Certificate of Origin

Data Sheets

Data Sheet - DUO92002

User Guide

Using Duolink in Multiwell Plates

Application Notes

Duolink Rehybridization Application Note
Visualizing DNA Methylation (5mC) on a Specific Genomic Locus (SEPTIN 9) in Individual Human Cancer Cells

Quotes and Ordering

Bulk Quote-Order Product

  1. 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.

  2. 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.

  3. 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 or view local office numbers.

  4. 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. 

  5. 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.

  6. 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;116:5297-305

  7. 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

  8. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Xiaofan Li et al.
PLoS pathogens, 13(3), e1006249-e1006249 (2017-03-02)
Trials to reintroduce chloroquine into regions of Africa where P. falciparum has regained susceptibility to chloroquine are underway. However, there are long-standing concerns about whether chloroquine increases lytic-replication of Epstein-Barr virus (EBV), thereby contributing to the development of endemic Burkitt
Birgitte B Olsen et al.
BMC molecular biology, 13, 7-7 (2012-03-13)
The DNA-dependent protein kinase (DNA-PK) is a nuclear complex composed of a large catalytic subunit (DNA-PKcs) and a heterodimeric DNA-targeting subunit Ku. DNA-PK is a major component of the non-homologous end-joining (NHEJ) repair mechanism, which is activated in the presence
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
M Aubele et al.
British journal of cancer, 103(5), 663-667 (2010-08-12)
Protein tyrosine kinase 6 (PTK6; breast tumour kinase) is overexpressed in up to 86% of the invasive breast cancers, and its association with the oncoprotein human epidermal growth factor receptor 2 (HER2) was shown in vitro by co-precipitation. Furthermore, expression
Jean Philippe Arnault et al.
Clinical cancer research : an official journal of the American Association for Cancer Research, 18(1), 263-272 (2011-11-19)
The emergence of skin tumors in patients treated with sorafenib or with more recent BRAF inhibitors is an intriguing and potentially serious event. We carried out a clinical, pathologic, and molecular study of skin lesions occurring in patients receiving sorafenib.
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Protocol for use of Duolink® PLA reagents for the detection of individual proteins, protein modifications, and protein-protein interactions within cell populations by flow cytometry.

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Duolink® PLA Applications

Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay

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