Description
Application
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 Starter 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 and the Duolink® flowPLA starter Kit. The flowPLA Starter Kits are available with 3 different fluorophores: Violet, Green, and FarRed. The flowPLA starter Kits contain all the necessary reagents reagents for 100 Duolink® PLA reactions, including a pair of PLA probes (Anti-Rabbit PLUS and Anti-Mouse MINUS) and all the 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, (rabbit and mouse-raised) 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
Duolink® flow PLA Mouse/Rabbit Starter Kit - Far Red has been used in the proximity ligase assay (PLA) to detect co-localization of human immune deficiency virus (HIV) protein, negative factor (Nef) with serine incorporator (SERINC3(iHA)) in human T-lymphoblastoid CEM A*02 cells.
Far Red Fluorescence Detection Reagents
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission.
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.
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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
Duolink® flowPLA Detection Kit – Far Red DUO94004-40TST
Duolink® In Situ PLA® Probe Anti-Mouse MINUS DUO92004-40TST
Duolink® In Situ PLA® Probe Anti-Rabbit PLUS DUO92002-40TST
Duolink® In Situ Wash Buffer, Brightfield (DUO82047-4L)
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
