MilliporeSigma
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DUO94005

Sigma-Aldrich

Duolink® flowPLA Detection Kit - Violet

Duolink® PLA kit for Flow Cytometry with Violet Detection

Synonym(s):
Anti-LAMS, Anti-PIERS, Anti-NPHS5

product line

Duolink®

technique(s)

flow cytometry: suitable
immunofluorescence: suitable
proximity ligation assay: suitable

fluorescence

λex 390 nm; λem 476 nm

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

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 100 RXN 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 registered trademark of Sigma-Aldrich Co. LLC

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Quotes and Ordering

Articles

How to Optimize Duolink® PLA for Flow Cytometry Detection

Considerations for proper experimental design, preparation and execution of the Duolink® PLA for flow cytometry protocol.

Troubleshooting Guide for Duolink® PLA for Flow Cytometry Detection

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.

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) with flow cytometry in a convenient kit, making the analysis of protein-protein interactions with flow cytometry readouts a reality.

Protocols

Duolink® PLA Flow Cytometry Protocol

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.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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