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DUO82047

Sigma-Aldrich

Duolink® In Situ Wash Buffer, Brightfield

Synonym(s):

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

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About This Item

UNSPSC Code:
12161703
NACRES:
NA.32

material

packing (powdered buffer pouches)

product line

Duolink®

technique(s)

proximity ligation assay: suitable

suitability

suitable for brightfield

storage temp.

20-25°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.

Follow the Duolink® In Situ Brightfield Protocol to use this product.

Visit our
Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.

To perform a complete Duolink® 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 HRP for brightfield detection.
Specificity
Duolink® In Situ Wash Buffer, Brightfield is used to wash specimen slides during Duolink® In Situ Brightfield procedures. See datasheet for more details.

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® 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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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

Legal Information

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

Storage Class Code

11 - Combustible Solids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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’.

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Agata Zieba et al.
Clinical chemistry, 56(1), 99-110 (2009-11-21)
The in situ proximity ligation assay (PLA) allows a protein or protein complex to be represented as an amplifiable DNA molecule. Recognition is mediated by proximity probes consisting of antibodies coupled with oligonucleotides. Upon dual binding of the proximity probes
Annabel Christ et al.
Developmental cell, 22(2), 268-278 (2012-02-22)
Sonic hedgehog (SHH) is a regulator of forebrain development that acts through its receptor, patched 1. However, little is known about cellular mechanisms at neurulation, whereby SHH from the prechordal plate governs specification of the rostral diencephalon ventral midline (RDVM)
Lei Zhu et al.
The Prostate, 73(2), 219-226 (2012-07-19)
PSA is the most useful prostate cancer marker. However, its levels are increased also in some non-malignant conditions. In circulation, the majority of PSA is complexed with protease inhibitors, including α(1) -antichymotrypsin (ACT). The proportion of the PSA-ACT complex is
Yoshihiro Akimoto et al.
Clinical proteomics, 8(1), 15-15 (2011-09-22)
The objective of the present study is to identify proteins that change in the extent of the modification with O-linked N-acetylglucosamine (O-GlcNAcylation) in the kidney from diabetic model Goto-Kakizaki (GK) rats, and to discuss the relation between O-GlcNAcylation and the
Timothy L Scott et al.
Molecular carcinogenesis, 56(2), 325-336 (2016-05-06)
Apurinic/apyrimidinic endonuclease 1 (APE1) is an essential protein crucial for repair of oxidized DNA damage not only in genomic DNA but also in mitochondrial DNA. Parkin, a tumor suppressor and Parkinson's disease (PD) associated gene, is an E3 ubiquitin ligase

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

Protocols

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.

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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