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Merck

DUO94004

Sigma-Aldrich

Duolink® flowPLA Detection Kit - FarRed

Duolink® PLA kit for Flow Cytometry with FarRed Detection

Synonym(e):

in situ Proximity Ligation Assay, Flowcytometry-PLA, Protein Protein Interaction Kit

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About This Item

UNSPSC-Code:
41105331
NACRES:
NA.32

Produktlinie

Duolink®

Methode(n)

flow cytometry: suitable
immunofluorescence: suitable
proximity ligation assay: suitable

Fluoreszenz

λex 644 nm; λem 669 nm

Eignung

suitable for fluorescence

Versandbedingung

dry ice

Lagertemp.

−20°C

Allgemeine Beschreibung

Duolink® flowPLA Detection Kit contains detection oligonucleotides with a fluorophore (lex = 644 nm/lem = 669 nm).

Spezifität

Far Red Fluorescence Detection Reagents
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission

Anwendung

Based on proximity ligation assay (PLA), the Duolink® PLA Technology allows for endogenous detection of protein interactions, post-translational modifications, and protein expression levels at the single molecule level in fixed cells.

Duolink® flowPLA Detection Kits will enable sensitive detection of proteins, protein-protein interactions, and protein modifications within cell populations by flow cytometry. To perform a Duolink® flowPLA experiment, you will need fixed, suspended cells, two primary antibodies that specifically recognize your proteins of interest, a pair of PLA probes (one 100RXN PLUS and one 100RXN MINUS), wash buffer, and a Duolink® flowPLA Detection Kit. The flowPLA Kits are available with 5 different fluorophores: Violet, Red, Green, Orange, or FarRed. The flowPLA Kits contain all the necessary reagents to perform the amplification and detection of bound PLA probes by flow cytometry. Analysis is carried out using standard flow cytometry assay equipment. User must provide a fixed cell suspension, primary antibodies, and corresponding PLA Probes.

Follow the Duolink® PLA Flow Cytometry Protocol to use this product.

Visit our Duolink® PLA Flow Cytometry page on how to run a Duolink® flow experiment, applications, troubleshooting, and more.

Application Note

Primary antibodies 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. Flow validated antibodies are recommended.

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

View full Duolink® product list
Duolink® flowPLA Detection Kit–FarRed has been used in in situ proximity ligation assay:
  • to detect interaction and complex formation between cellular retinoic acid binding protein 1 (Crabp1) and the components of rapidly accelerated fibrosarcoma (Raf) kinase- MAPK-Erk kinase (Mek) signaling pathway
  • to study protein interaction in human cultured MOLT-4 cells and HeLa cells
  • to visualize Beclin-1 protein interaction with 14-3-3t in neurons
  • to study protein interactions in graft endothelial cells 

Leistungsmerkmale und Vorteile

  • Analyze protein protein interactions with flow cytometry readout
  • Analyze cell populations with Proximity Ligation Assay
  • Increased sensitivity due to rolling circle amplification for low abundant targets
  • No overexpression or genetic manipulation required
  • Relative quantification possible
  • Works with any flow cytometer instrumentation
  • Easy to follow flexible protocol
  • Publication-ready results

Komponenten

This product is comprised of the following:
  • 5x Detection Solution - FarRed (DUO84004)
  • 5x Ligation Buffer (DUO82009)
  • 5x Amplification Buffer (DUO82050)
  • Ligase (1U/μL)
  • Polymerase (10U/μL)

See datasheet for more information.

Rechtliche Hinweise

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

Piktogramme

Health hazard

Signalwort

Danger

H-Sätze

Gefahreneinstufungen

Resp. Sens. 1

Lagerklassenschlüssel

10 - Combustible liquids


Analysenzertifikate (COA)

Suchen Sie nach Analysenzertifikate (COA), indem Sie die Lot-/Chargennummer des Produkts eingeben. Lot- und Chargennummern sind auf dem Produktetikett hinter den Wörtern ‘Lot’ oder ‘Batch’ (Lot oder Charge) zu finden.

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In der Dokumentenbibliothek finden Sie die Dokumentation zu den Produkten, die Sie kürzlich erworben haben.

Die Dokumentenbibliothek aufrufen

Adriana Cassaro et al.
Hematological oncology, 39(3), 364-379 (2021-01-27)
Wnt/Fzd signaling has been implicated in hematopoietic stem cell maintenance and in acute leukemia establishment. In our previous work, we described a recurrent rearrangement involving the WNT10B locus (WNT10BR ), characterized by the expression of WNT10BIVS1 transcript variant, in acute
Tyler J Burns et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 91(2), 180-189 (2017-01-18)
To quantify visual and spatial information in single cells with a throughput of thousands of cells per second, we developed Subcellular Localization Assay (SLA). This adaptation of Proximity Ligation Assay expands the capabilities of flow cytometry to include data relating
Sofie Selmer Andersen et al.
Cytokine, 64(1), 54-57 (2013-06-04)
Many cytokine receptors are cell surface proteins that promiscuously combine to form active signalling homo- or heterodimers. Thus, receptor chain dimerization can be viewed as a direct measure of a high probability of intracellular signalling by specific cytokines. Proximity ligation
Sung Wook Park et al.
Scientific reports, 9(1), 10929-10929 (2019-07-31)
The rapidly accelerated fibrosarcoma (Raf) kinase is canonically activated by growth factors that regulate multiple cellular processes. In this kinase cascade Raf activation ultimately results in extracellular regulated kinase 1/2 (Erk1/2) activation, which requires Ras binding to the Ras binding
Karl-Johan Leuchowius et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 75(10), 833-839 (2009-08-04)
Interactions between members of the epidermal growth factor receptor (EGFR) family mediates cellular responses to ligand stimulation. Measurement of these interactions could provide important information and may prove useful as prognostic markers in malignancy. Therefore, to develop methods to study

Artikel

Considerations for proper experimental design, preparation and execution of the Duolink® PLA for flow cytometry protocol.

Duolink® PLA kit enhances flow cytometry for detecting protein interactions accurately.

General tips and tricks for proper experiment execution, aid in identifying potential problems, and provide solutions to ensure a successful Duolink® PLA experiment for flow cytometry.

Erfahren Sie, wie die Proximity Ligation Assay-Technologie funktioniert und wie durch das Kontrollkit für die Protein-Protein-Interaktion der In-situ-Nachweis der EGF-induzierten EGFR-HER2-Dimerisierung bestätigt werden kann.

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