Not available outside of the UK & Ireland.

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 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 NotePrimary 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^® projectsView 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 pathwayto study protein interaction in human cultured MOLT-4 cells and HeLa cellsto visualize Beclin-1 protein interaction with 14-3-3tin neuronsto study protein interactions in graft endothelial cells

Components

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.

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

General description

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

Legal Information

Duolink is a registered trademark of Sigma-Aldrich Co. LLC

PLA is a registered trademark of Sigma-Aldrich Co. LLC

Specificity

Far Red Fluorescence Detection ReagentsUse appropriate laser for λex 644 nm excitationUse appropriate filter for λem 669 nm emission