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Key Documents

DUO92003

Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Goat PLUS

Synonyme(s) :

in situ Proximity Ligation Assay Kit, Protein Protein Interaction Kit

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About This Item

Code UNSPSC :
12352203
Nomenclature NACRES :
NA.32

Source biologique

donkey (polyclonal)

Niveau de qualité

Forme d'anticorps

affinity purified immunoglobulin (secondary antibody)

Type de produit anticorps

primary antibodies

Gamme de produits

Duolink®

Espèces réactives

goat

Technique(s)

immunofluorescence: suitable
proximity ligation assay: suitable

Adéquation

suitable for brightfield
suitable for fluorescence

Conditions d'expédition

wet ice

Température de stockage

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-Goat reacts with whole molecule goat IgG and the light chains of other goat immunoglobulin?s. The PLA probe anti-Goat may cross-react with sheep antibodies, but has minimal cross reactivity with chicken, guinea pig, Syrian hamster, horse, human, mouse, rabbit, and rat 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.

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View full Duolink® product list
Duolink® In Situ PLA® Probe Anti-Goat PLUS has been used in in situ proximity ligation assay in primary hippocampal neuronal cultures, mesenteric arteries and human malignant melanoma cell line(MEL624).

Caractéristiques et avantages

  • 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

Composants

This product is comprised of the following:
  • 5x PLA Probe Anti-Goat PLUS - Donkey anti-goat 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.

Informations légales

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

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Code de la classe de stockage

10 - Combustible liquids


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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

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Miyamoto T, et al.
The Journal of Biological Chemistry, 291(4), 1719-1734 (2016)
Targeting ER stress-induced autophagy overcomes BRAF inhibitor resistance in melanoma
Ma XH, et al.
The Journal of Clinical Investigation, 124(3), 1406-1417 (2014)
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)
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Articles

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

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

Protocoles

Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.

Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.

Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.

Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.

Contenu apparenté

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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