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DUO92012

Sigma-Aldrich

Duolink® In Situ Detection Reagents Brightfield

Synonim(y):

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

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About This Item

Kod UNSPSC:
12352200

linia produktu

Duolink®

opakowanie

pkg of Box A (-20°C)
pkg of Box B (2-8°C)

metody

proximity ligation assay: suitable

przydatność

suitable for brightfield

temp. przechowywania

−20°C

Powiązane kategorie

Zastosowanie

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® 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 HRP for brightfield detection.
Specificity
Brightfield detection reagents are used for samples with significant autofluorescence.

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.

Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects

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Cechy i korzyści

  • 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

Komponenty

This product is comprised of the following:
Box A:
  • 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
  • 1x Ligase (1 unit/μL)
  • 5x Amplification - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase
  • 5x Detection Brightfield - Contains oligonucleotide probes labeled with horseradish peroxidase (HRP) that hybridize to the RCA product
  • 1x Polymerase (10 units/μL)

Box B:
  • 1x Hydrogen Peroxide - Contains 0.3% hydrogen peroxide
  • Substrate Reagents A-D - Contains all substrate components needed for HRP enzymatic reaction
  • 1x Nuclear Stain - Contains Mayer′s hematoxylin solution for cell nuclei staining
See datasheet for more information.

Not included in Detection kit:

Primary antibodies, PLA probes, wash buffers, mounting medium

Inne uwagi

Storage/Stability
Store Box A and all of its components at -20°C. The enzymes should be kept cold (-20°C) at all times; Use a freezing block when removing them from the freezer.
Store Box B and all of its components at 2-8°C.

Informacje prawne

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany
This page may contain text that has been machine translated.

Hasło ostrzegawcze

Danger

Klasyfikacja zagrożeń

Acute Tox. 3 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3 - Eye Dam. 1 - Flam. Liq. 2 - Muta. 2 - Skin Irrit. 2 - Skin Sens. 1

Kod klasy składowania

3 - Flammable liquids

Klasa zagrożenia wodnego (WGK)

WGK 3

Temperatura zapłonu (°F)

42.8 °F

Temperatura zapłonu (°C)

6 °C

Certyfikaty analizy (CoA)

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Odwiedź Bibliotekę dokumentów

Salvador Sierra et al.
Brain structure & function, 220(5), 2721-2738 (2014-06-29)
Although type 1 cannabinoid receptors (CB1Rs) are expressed abundantly throughout the brain, the presence of type 2 cannabinoid receptors (CB2Rs) in neurons is still somewhat controversial. Taking advantage of newly designed CB1R and CB2R mRNA riboprobes, we demonstrate by PCR
Zhiwei Ang et al.
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 32(1), 289-303 (2017-09-09)
Free fatty acid receptors 2 and 3 (FFAR2/FFA2/GPR43 and FFAR3/FFA3/GPR41) are mammalian receptors for gut microbiota-derived short-chain fatty acids (SCFAs). These receptors are promising drug targets for obesity, colitis, colon cancer, asthma, and arthritis. Here, we demonstrate that FFAR2 and
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
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Duolink® References

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

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Duolink® PLA Troubleshooting Guide

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

Protokoły

Duolink® PLA Brightfield Protocol

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.

Nasz zespół naukowców ma doświadczenie we wszystkich obszarach badań, w tym w naukach przyrodniczych, materiałoznawstwie, syntezie chemicznej, chromatografii, analityce i wielu innych dziedzinach.

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