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DUO94004

Sigma-Aldrich

Duolink® flowPLA Detection Kit - FarRed

Duolink® PLA kit for Flow Cytometry with FarRed Detection

Synonym(s):

in situ Proximity Ligation Assay, Flowcytometry-PLA, Protein Protein Interaction Kit

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About This Item

UNSPSC Code:
41105331
NACRES:
NA.32

product line

Duolink®

technique(s)

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

fluorescence

λex 644 nm; λem 669 nm

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

General description

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

Specificity

Far Red Fluorescence Detection Reagents
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission

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 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

View 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 pathway
  • to study protein interaction in human cultured MOLT-4 cells and HeLa cells
  • to visualize Beclin-1 protein interaction with 14-3-3t in neurons
  • to study protein interactions in graft endothelial cells 

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 - FarRed (DUO84004)
  • 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

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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