Description
Application
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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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.
Biochem/physiol Actions
Orange Fluorescence Detection Reagents
Use appropriate laser for λex 554 nm excitation
Use appropriate filter for λem 576 nm emission
Features and Benefits
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
Other Notes
This product is comprised of the following:
5x Detection Solution - Orange (DUO84003)
5x Ligation Buffer (DUO82009)
5x Amplification Buffer (DUO82050)
Ligase (1U/μL)
Polymerase (10U/μL)
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
