DUO94004
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
About This Item
Empfohlene Produkte
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
Spezifität
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission
Anwendung
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.
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View full Duolink® product list
- 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
- 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
Signalwort
Danger
H-Sätze
P-Sätze
Gefahreneinstufungen
Resp. Sens. 1
Lagerklassenschlüssel
10 - Combustible liquids
Analysenzertifikate (COA)
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Artikel
Considerations for proper experimental design, preparation and execution of the Duolink® PLA for flow cytometry protocol.
Traditional flow cytometry has been limited in the ability to detect protein-protein interactions and low abundant proteins events — until now. We have combined Duolink® Proximity Ligation Assay (PLA) with flow cytometry in a convenient kit, making the analysis of protein-protein interactions with flow cytometry readouts a reality.
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.
Protokolle
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.
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