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DUO94104

Sigma-Aldrich

Duolink® flowPLA Mouse/Rabbit Starter Kit - Far Red

Duolink® PLA kit for Flow Cytometry with Far Red Detection and Mouse/Rabbit probes

Synonym(s):

in situ Proximity Ligation Assay Kit, Protein Protein Interaction Kit

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About This Item

UNSPSC Code:
41116158
NACRES:
NA.41

Quality Level

200

technique(s)

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

fluorescence

λex 644 nm; λem 669 nm

suitability

suitable for fluorescence

storage temp.

−20°C

Related Categories

Specificity

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.

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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.

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

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.

Principle

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.

Legal Information

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany
Plexiglas is a registered trademark of Arkema

pictograms

Health hazardEnvironment
GHS08,GHS09

signalword

Danger

Hazard Classifications

Aquatic Chronic 2 - Resp. Sens. 1 - Skin Sens. 1

Storage Class

10 - Combustible liquids

Certificates of Analysis (COA)

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Steven W Jin et al.
PLoS pathogens, 16(9), e1008813-e1008813 (2020-09-15)
HIV Nef counteracts cellular host restriction factors SERINC3 and SERINC5, but our understanding of how naturally occurring global Nef sequence diversity impacts these activities is limited. Here, we quantify SERINC3 and SERINC5 internalization function for 339 Nef clones, representing the

Articles

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

Duolink® PLA kit enhances flow cytometry for detecting protein interactions accurately.

How Proximity Ligation Assays (PLA) Work

Learn how Proximity Ligation Assay technology works and how the protein-protein interaction control kit can confirm in situ detection of EGF-induced EGFR-HER2 dimerization.

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