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DUO94005 Sigma-Aldrich

Duolink flowPLA Detection Kit - Violet

Duolink PLA kit for Flow Cytometry with Violet Detection

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Properties

Related Categories Duolink PLA Technology, Duolink flowPLA Detection Kits, Molecular Biology, Protein Interaction, Proteomics More...
product line   Duolink
application(s)   flow cytometry: suitable
  immunofluorescence: suitable
  proximity ligation assay: suitable
fluorescence   λex 390 nm; λem 476 nm (Violet)
suitability   suitable for fluorescence
shipped in   dry ice
storage temp.   −20°C

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 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 PLUS and one MINUS), wash buffer and a Duolink flowPLA Detection Kit. The flowPLA Kits are available with 5 different fluorophores: Violet, Green, Orange, Red, 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.

Specificity

Violet Fluorescence Detection Reagents
Use appropriate laser for λex 390 nm excitation
Use appropriate filter for λem 476 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.

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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

Components

This product is comprised of the following:
• 5x Detection Solution - Violet (DUO84005)
• 5x Ligation Buffer (DUO82009)
• 5x Amplification Buffer (DUO82050)
• Ligase (1U/μL)
• Polymerase (10U/μL)

See datasheet for more information.

Legal Information

Duolink is a trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Duolink® PLA Flow Cytometry Kits

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...
Keywords: Amplification, Flow cytometry, Peptide synthesis

How to Optimize Duolink® PLA for Flow Cytometry Detection

Adaptation of Duolink® in situ proximity ligation assay (PLA) into flow cytometry workflows allows quantitative detection of protein-protein interactions, protein post-translational modifications, an...
Keywords: Bacterial conjugations, Buffers, Cell dissociation, Centrifugation, Detection methods, Detergents, Flow cytometry, Gene expression, Growth factors, Immunofluorescence, Microscopy, Peptide synthesis, Phosphorylations, Post translational modifications, Sample preparations, Titrations

Troubleshooting Guide for Duolink® PLA for Flow Cytometry Detection

The combination of Duolink® PLA technology and flow cytometry allows quantitative detection of protein-protein interactions, post-translational modifications, and protein expression with greater stat...
Keywords: Amplification, Anticoagulants, Buffers, Centrifugation, Detection methods, Detergents, Flow cytometry, Gene expression, Immunofluorescence, Microscopy, Peptide synthesis, Titrations

Protocols

Duolink® PLA Flow Cytometry Protocol

This protocol describes the use of Duolink® PLA reagents for the detection of individual proteins, protein modifications, and protein-protein interactions within cell populations by flow cytometry.
Keywords: Amplification, Centrifugation, Flow cytometry, Microscopy, Peptide synthesis

Duolink® PLA Product Selection Guide

Getting started with Duolink® PLA is simple. The protocols are very similar to that of IF, IHC, and flow cytometry, however Duolink® PLA offers distinct advantages over these and other traditional me...
Keywords: Amplification, Bacterial conjugations, Buffers, Flow cytometry, Fluorescent microscopy, Gene expression, Immunohistochemistry, Microscopy, Peptide synthesis

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