质量水平
技术
flow cytometry: suitable
immunofluorescence: suitable
proximity ligation assay: suitable
荧光
λex 390 nm; λem 476 nm
适用性
suitable for fluorescence
储存温度
−20°C
特异性
Use appropriate laser for λex 390 nm excitation
Use appropriate filter for λem 476 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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应用
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.
特点和优势
- 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
组分
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.
法律信息
PLA is a registered trademark of Sigma-Aldrich Co. LLC
警示用语：
Danger
危险声明
危险分类
Aquatic Chronic 2 - Resp. Sens. 1 - Skin Sens. 1
储存分类代码
10 - Combustible liquids
法规信息
商品
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。
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 kit enhances flow cytometry for detecting protein interactions accurately.
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.
实验方案
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。
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