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DUO92001

Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Mouse PLUS

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Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent

biological source

donkey (polyclonal)

Quality Level

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody product type

primary antibodies

product line

Duolink®

species reactivity

mouse

technique(s)

immunofluorescence: suitable
proximity ligation assay: suitable

suitability

suitable for brightfield
suitable for fluorescence

shipped in

wet ice

storage temp.

2-8°C

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This Item
DUO92002DUO92005DUO92003
antibody form

affinity purified immunoglobulin (secondary antibody)

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody form

affinity purified immunoglobulin (secondary antibody)

biological source

donkey (polyclonal)

biological source

donkey (polyclonal)

biological source

donkey (polyclonal)

biological source

donkey (polyclonal)

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

product line

Duolink®

product line

Duolink®

product line

Duolink®

product line

Duolink®

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

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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

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.

Legal Information

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany

Pictograms

Exclamation markEnvironment

Signal Word

Warning

Hazard Statements

Hazard Classifications

Aquatic Chronic 2 - Skin Sens. 1

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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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
Xiaofan Li et al.
Journal of virology, 93(17) (2019-06-14)
Herpesviruses are ubiquitous, and infection by some, like Epstein-Barr virus (EBV), is nearly universal. To persist, EBV must periodically switch from a latent to a replicative/lytic phase. This productive phase is responsible for most herpesvirus-associated diseases. EBV encodes a latency-to-lytic
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
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
John J Peluso et al.
Endocrinology, 153(8), 3929-3939 (2012-06-22)
Progesterone (P4) receptor membrane component (PGRMC)1 is detected as a 22-kDa band as well as higher molecular mass bands (>50 kDa) in spontaneously immortalized granulosa cells. That these higher molecular mass bands represent PGRMC1 is supported by the findings that

Articles

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

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

Protocols

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.

Related Content

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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