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DUO92003

Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Goat PLUS

Synonym(s):

in situ Proximity Ligation Assay Kit, Protein Protein Interaction Kit

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.32

biological source

donkey (polyclonal)

Quality Level

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody product type

primary antibodies

product line

Duolink®

species reactivity

goat

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-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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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).

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-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.

Legal Information

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

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Storage Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

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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)
Increasing the receptor tyrosine kinase EphB2 prevents amyloid-beta-induced depletion of cell surface glutamate receptors by a mechanism that requires the PDZ-binding motif of EphB2 and neuronal activity
Miyamoto T, et al.
The Journal of Biological Chemistry, 291(4), 1719-1734 (2016)
CaMKII regulates intracellular Ca2+ dynamics in native endothelial cells
Toussaint F, et al.
Cell Calcium, 58(3), 275-285 (2015)
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
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

Articles

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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

Protocols

This protocol describes the use of Duolink® PLA reagents for the brightfield detection, visualization, and quantification of individual proteins, protein modifications, and protein interactions in tissue and cell samples.

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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