Skip to Content
MilliporeSigma
All Photos(2)

Key Documents

View All Documentation

DUO92005

Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Rabbit MINUS

Synonym(s):

in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent

Sign Into View Organizational & Contract Pricing
All Photos(2)

About This Item

UNSPSC Code:
12352203
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

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, mouse, rat, and sheep serum proteins. A PLUS 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

View full Duolink® product list

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 MINUS - Donkey anti-rabbit secondary antibody conjugated to oligonucleotide MINUS
  • 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 Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany

Not finding the right product?  

Try our Product Selector Tool.

Storage Class

10 - Combustible liquids

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

Already Own This Product?

Find documentation for the products that you have recently purchased in the Document Library.

Visit the Document Library

Roberto Di Maio et al.
Science translational medicine, 10(451) (2018-07-27)
Missense mutations in leucine-rich repeat kinase 2 (LRRK2) cause familial Parkinson's disease (PD). However, a potential role of wild-type LRRK2 in idiopathic PD (iPD) remains unclear. Here, we developed proximity ligation assays to assess Ser1292 phosphorylation of LRRK2 and, separately
Hyun Jik Lee et al.
Cell death and differentiation, 26(9), 1716-1734 (2018-11-23)
Hypoxia inducible factor 1α (HIF1α) is a master regulator leading to metabolic adaptation, an essential physiological process to maintain the survival of stem cells under hypoxia. However, it is poorly understood how HIF1α translocates into the nucleus in stem cells
Kan V Lu et al.
Cancer cell, 22(1), 21-35 (2012-07-14)
Inhibition of VEGF signaling leads to a proinvasive phenotype in mouse models of glioblastoma multiforme (GBM) and in a subset of GBM patients treated with bevacizumab. Here, we demonstrate that vascular endothelial growth factor (VEGF) directly and negatively regulates tumor
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
View All Related Papers

Articles

How to Optimize the Duolink® Proximity Ligation Assay

Things to consider for preparation, setup and execution of the Duolink® assay protocol

Duolink® References

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

Duolink® PLA Troubleshooting Guide

Support information including tips and tricks, frequently asked questions, and basic troubleshooting.

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.

Contact Technical Service