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DUO94004

Sigma-Aldrich

Duolink® flowPLA 检测试剂盒- FarRed

Duolink® PLA kit for Flow Cytometry with FarRed Detection

同義詞:

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

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About This Item

分類程式碼代碼:
41105331
NACRES:
NA.32
暫時無法取得訂價和供貨情況

產品線

Duolink®

技術

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

螢光

λex 644 nm; λem 669 nm

適合性

suitable for fluorescence

運輸包裝

dry ice

儲存溫度

−20°C

相關類別

一般說明

Duolink® flowPLA检测试剂盒包含含有荧光团的检测寡核苷酸(lex = 644 nm/lem = 669 nm)。

特異性

远红荧光检测试剂
使用合适的激光器进行λex 644 nm激发
使用合适的滤光片进行λem 669 nm发射

應用

Duolink® flowPLA检测试剂盒–FarRed已用于原位邻位连接试验：
  • 检测细胞维甲酸结合蛋白1 (Crabp1)与快速加速纤维肉瘤(Raf)激酶- MAPK-Erk激酶(Mek)信号通路组分之间的相互作用和复合物形成[1]
  • 研究人类培养的MOLT-4细胞和HeLa细胞中的蛋白质相互作用[2]
  • 以可视化Beclin-1蛋白与14-3-3t在神经元中的相互作用[3]
  • 研究移植内皮细胞中蛋白质的相互作用[4] 

基于邻位连接技术（PLA），Duolink® PLA可实现在单分子水平上内源性检测固定细胞中的蛋白质相互作用、翻译后修饰和蛋白表达水平。

Duolink® flowPLA检测试剂盒可通过流式细胞术灵敏地检测细胞群内蛋白质、蛋白质-蛋白质相互作用和蛋白质修饰。Duolink® flowPLA实验需要固定的悬浮细胞、两种可特异性识别目标蛋白的一抗、一对PLA探针（一个PLUS和一个MINUS）、洗涤缓冲液和Duolink® flowPLA检测试剂盒。flowPLA试剂盒具有5种不同荧光染料：紫色、红色、绿色、橙色或远红外。flowPLA试剂盒包含流式细胞术对结合的PLA探针进行扩增和检测所需的所有试剂。使用标准流式细胞术分析设备进行分析。使用者必须提供固定的细胞悬液、一抗和相应的OLA探针。

遵循Duolink® PLA流式细胞术方案使用本产品。

请访问我们的Duolink® PLA流式细胞术页面，了解如何运行Duolink®流式实验、应用程序、故障排除等。

应用说明

需要一抗。采用标准免疫荧光（IF）、免疫组化（IHC）或免疫细胞化学（ICC）测定法检测一抗（IgG类，单克隆或多克隆），确定最佳的固定、封闭和滴度条件。推荐经流式验证的抗体。

请让我们助您完成工作，了解有关我们的定制服务计划的更多信息，加速您的Duolink®项目

查看完整的Duolink®产品列表

特點和優勢

  • 采用流式细胞仪分析蛋白质与蛋白质的相互作用
  • 邻近连接法分析细胞群
  • 通过滚环扩增提高对低丰度靶标的灵敏度
  • 不需要过表达或基因操纵
  • 可进行相对量化
  • 可与任何流式细胞仪配合使用
  • 易于遵循灵活方案
  • 结果可直接发表

成分

本品包括以下组分：
  • 5x检测溶液-FarRed（DUO84004）
  • 5x连接缓冲液（DUO82009）
  • 5x扩增缓冲液（DUO82050）
  • 连接酶（1U/μL）
  • 聚合酶（10U/μL）

有关更多信息，参见数据表。

法律資訊

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

象形圖

Health hazard
GHS08

訊號詞

Danger

危險聲明

H334

防範說明

P261 - P284 - P501

危險分類

Resp. Sens. 1

儲存類別代碼

10 - Combustible liquids

從最近期的版本中選擇一個：

分析證明 (COA)

Lot/Batch Number
SLCR4421
SLCM9271
SLCL5872
SLCJ9163
SLCD9319

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如果您需要一個特定的版本，您可以透過批號來尋找特定憑證。

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您可以在文件庫中找到最近購買的產品相關文件。

存取文件庫

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
Sung Wook Park et al.
Scientific reports, 9(1), 10929-10929 (2019-07-31)
The rapidly accelerated fibrosarcoma (Raf) kinase is canonically activated by growth factors that regulate multiple cellular processes. In this kinase cascade Raf activation ultimately results in extracellular regulated kinase 1/2 (Erk1/2) activation, which requires Ras binding to the Ras binding
Adriana Cassaro et al.
Hematological oncology, 39(3), 364-379 (2021-01-27)
Wnt/Fzd signaling has been implicated in hematopoietic stem cell maintenance and in acute leukemia establishment. In our previous work, we described a recurrent rearrangement involving the WNT10B locus (WNT10BR ), characterized by the expression of WNT10BIVS1 transcript variant, in acute
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
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文章

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.

Duolink® PLA流式蛋白检测试剂盒

了解有关将邻位连接技术 (PLA) 与流式细胞术相结合的新试剂盒的更多信息，实现了蛋白互作流式分析。

Duolink® PLA Flow Cytometry Kits

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

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.

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條款

Duolink®PLA流式检测方案

本实验方案描述如何使用Duolink® PLA邻位连接试剂，通过流式细胞术检测细胞群内的单个蛋白质、蛋白修饰和蛋白相互。

Questions

1–3 of 3 Questions  

  1. Is it possible to just buy the 5x Detection Solution separately?

    1 answer

    1. Unfortunately, the kit components are not sold separately.

      Helpful?

  2. Can I use this kit in conjunction with staining for other markers? If so, would you recommend staining with the PLA kit first, followed by staining with other primary-conjugated antibodies?

    1 answer

    1. It is possible to counterstain after the Duolink in situ amplification. It is recommended to apply the counterstaining protocol after the completion of the Amplification step in section 7.3, step 5 of the Duolink In Situ Fluorescence User Manual.

      Please see the protocol below:
      https://www.sigmaaldrich.com/technical-documents/protocol/protein-biology/protein-and-nucleic-acid-interactions/duolink-counterstain

      Helpful?

  3. Hello, can I also dilute the 5 x detection stock in something else than high purity water? Maybe PBS?

    1 answer

    1. It is not recommended to use any other solvent to dilute the 5x detection stock due to the sensitivity of this reaction. The Duolink kits have been optimized for use under the conditions described. Deviation from the protocol may not offer the best results. Please see the link below to view the full product information for the Duolink PLAFlow kits:
      https://www.sigmaaldrich.com/technical-documents/technical-article/protein-biology/protein-and-nucleic-acid-interactions/duolink-pla-flow-cytometry-kits

      Helpful?

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