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Key Documents

DUO96001

Sigma-Aldrich

Duolink® flowPLA Multicolor Reagent Pack

Duolink® PLA kit for Flow Cytometry Multiplexing

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About This Item

Code UNSPSC :
41116158
Nomenclature NACRES :
NA.32

Niveau de qualité

Gamme de produits

Duolink®

Technique(s)

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

Conditions d'expédition

dry ice

Température de stockage

−20°C

Spécificité

Fluorescence detection reagents are often used with FITC, Cy®3,Texas Red® or Cy®5 filter. The filter chosen is dependent on the color of the Multicolor Probemaker kit selected.

Duolink® flowPLA Multicolor Reagent Pack contains all the necessary reagents to perform the amplification and detection of bound custom PLA probes generated using Duolink® PLA Multicolor Probemaker Kits. Sufficient components are provided for approximately 40 tests at 100 μL reaction volume.

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® Multicolor flow PLA Reagent Pack Kits used with Duolink® PLA Multicolor Probemaker Kits will enable Multiplexing of up to 4 protein events with 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 and permeabilized suspended cells, and at least two primary antibodies that specifically recognize your proteins of interest. Analysis is carried out using standard flow cytometry assay equipment. User must provide a fixed cell suspension and primary antibodies.

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. Duolink® in situ reagents are suitable for use on fixed cells, cytospin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). Flow validated antibodies are recommended. It is also highly recommended that users purchase our Duolink(insert registration symbol) Compensation Beads (make title a hyperlink to beads products) to inform experimental design, instrument settings, and assist with compensation parameters.

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Caractéristiques et avantages

  • Visualize and detect multiple proteins events in one experiment
  • No overexpression or genetic manipulation required
  • High specificity (fewer false positives)
  • Detection of low abundant proteins due to rolling circle amplification
  • Relative quantification possible
  • No special equipment needed
  • Faster and more simple than FRET
  • Publication-ready results

Composants

This product is comprised of the following:
  • 5x Multicolor PLA Ligation Buffer (DUO86002)
  • 5x Amplification Buffer (DUO82050)
  • 5x Multicolor FlowPLA Detection Buffer (DUO86003)
  • 1x Ligase (DUO82025)
  • 1x Polymerase (DUO82026)
  • 1x Blocking Solution (DUO82007)
  • Duolink Probemaker PLA Probe Diluent (DUO82036)

See datasheet for more information.

Informations légales

Cy is a registered trademark of Cytiva
Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
Texas Red is a registered trademark of Life Technologies

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Mention d'avertissement

Warning

Mentions de danger

Classification des risques

Aquatic Chronic 2 - Skin Sens. 1

Code de la classe de stockage

10 - Combustible liquids


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