DUO94004
Duolink® flowPLA Detection Kit - FarRed
Duolink® PLA kit for Flow Cytometry with FarRed Detection
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About This Item
NACRES:
NA.32
UNSPSC Code:
41105331
Fluorescence:
λex 644 nm; λem 669 nm
Technique(s):
flow cytometry: suitable, immunofluorescence: suitable, proximity ligation assay: suitable
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).
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 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
Biochem/physiol Actions
Far Red Fluorescence Detection Reagents
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission
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
Other Notes
This product is comprised of the following:
See datasheet for more information.
- 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
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Danger
Storage Class
10 - Combustible liquids
wgk
WGK 3
hcodes
Hazard Classifications
Aquatic Chronic 2 - ED ENV 1 - Resp. Sens. 1
Regulatory Listings
Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.
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fsl
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ishl_indicated
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ishl_notified
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Tyler J Burns et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 91(2), 180-189 (2017-01-18)
To quantify visual and spatial information in single cells with a throughput of thousands of cells per second, we developed Subcellular Localization Assay (SLA). This adaptation of Proximity Ligation Assay expands the capabilities of flow cytometry to include data relating
Sofie Selmer Andersen et al.
Cytokine, 64(1), 54-57 (2013-06-04)
Many cytokine receptors are cell surface proteins that promiscuously combine to form active signalling homo- or heterodimers. Thus, receptor chain dimerization can be viewed as a direct measure of a high probability of intracellular signalling by specific cytokines. Proximity ligation
Karl-Johan Leuchowius et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 75(10), 833-839 (2009-08-04)
Interactions between members of the epidermal growth factor receptor (EGFR) family mediates cellular responses to ligand stimulation. Measurement of these interactions could provide important information and may prove useful as prognostic markers in malignancy. Therefore, to develop methods to study
Teng-Jian Zhou et al.
Theranostics, 7(5), 1389-1406 (2017-04-25)
Cancer stem cells (CSCs) are a small subset of malignant cells, possessing stemness, with strong tumorigenic capability, conferring resistance to therapy and leading to the relapse of nasopharyngeal carcinoma (NPC). Our previous study suggested that cyclooxygenase-2 (COX-2) would be a
Valerie Le Sage et al.
Virology, 502, 73-83 (2016-12-26)
Stress granules (SGs) are dynamic cytoplasmic aggregates of translationally silenced mRNAs that assemble in response to environmental stress. SGs appear to play an important role in antiviral innate immunity and many viruses have evolved to block or subvert SGs components
Articles
フローサイトメトリープロトコルのための、適切な実験デザイン、Duolink® PLAの準備と実施する際の検討事項
フローサイトメトリーのためのDuolink™ PLA実験で成果を上げるために、正しく実験を行うための一般的なヒントやコツ、問題を特定するための有用な情報と解決策を提供します。
フローサイトメトリーの測定値でタンパク質間相互作用と分析できる、近接ライゲーションアッセイ（PLA）とフローサイトメトリーを組み合わせた新しいキットについてご説明します。
近接ライゲーションアッセイ技術の仕組みと、EGFによるEGFR-HER2二量体化のin situ検出を確認できるタンパク質間相互作用コントロールキットの仕組みについてご紹介します
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