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Merck
CN

DUO82049

Duolink® In Situ Wash Buffers, Fluorescence

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About This Item

NACRES:
NA.32
UNSPSC Code:
12161703
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material

packing (powdered buffer pouches)

Quality Level

product line

Duolink®

technique(s)

proximity ligation assay: suitable

suitability

suitable for fluorescence

storage temp.

20-25°C

Application

Duolink® PLA 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).
Duolink®proximity ligation assay(PLA®) allows for endogenous detection of protein interactions, post translational modifications, and protein expression levels at the single molecule level in fixed cells and tissue samples.

Use the Duolink® In Situ Fluorescence Protocol for this product. A set of short instructionsis also available.

Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.

To perform a complete Duolink® PLA in situ experiment you will need two primary antibodies (PLA, IHC, ICC or IF validated) that recognize two target epitopes. Other necessary reagents include a pair of PLA probes from different species (one PLUS and one MINUS), detection reagents, wash buffers, and mounting medium. Note that the primary antibodies must come from the same species as the Duolink® PLA probes. Analysis is carried out using standard immunofluorescence assay equipment.
Specificity
Duolink® In Situ fluorescence applications use two wash buffers. Wash Buffer A is used after the PLA Probe incubation step and Wash Buffer B is used after incubation with the amplification reagents. See datasheet for more information.

Application Note
Two primary antibodies raised in different species 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®PLA 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). No minimum number of cells is required.

Linkage
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects

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Features and Benefits

  • No overexpression or genetic manipulation required
  • High specificity (fewer false positives)
  • Single molecule sensitivity due to rolling circle amplification
  • Relative quantification possible
  • No special equipment needed
  • Quicker and simpler than FRET
  • Increased accuracy compared to co-IP
  • Publication-ready results

Legal Information

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


Storage Class

10 - Combustible liquids

wgk

WGK 3



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Protocols

使用荧光显微镜和多色试剂同时检测、可视化和定量一个组织或细胞样本中多达4种的蛋白、蛋白修饰或蛋白互作。

本页详细介绍了Duolink®原位精简实验方案

This protocol describes how to perform immunofluorescent detection of proteins in cells and tissue.

View All Protocols

Articles

本实验方案描述了Duolink® PLA试剂用于组织和细胞样品中个体蛋白质、蛋白质修饰和蛋白质相互作用的免疫荧光检测、可视化和定量。

包括建议和技巧、常见问题解答和基本故障排除的支持信息。

正确地准备、设置和执行 Duolink® PLA 实验方案需要考虑的因素。

Related Content

利用邻位连接技术对蛋白质互作、翻译后修饰和低表达蛋白进行检测、定量和成像应用。

View an automated protocol for Duolink® assays on the AAW™ automated assay workstation and see results comparing manual vs automated runs.


Increasing the Receptor Tyrosine Kinase EphB2 Prevents Amyloid-?-induced Depletion of Cell Surface Glutamate Receptors by a Mechanism That Requires the PDZ-binding Motif of EphB2 and Neuronal Activity
Miyamoto T, et al.
The Journal of Biological Chemistry (2015)
Detection of Receptor Heteromerization Using In Situ Proximity Ligation Assay
Gomes I, et al.
Current Protocols in Pharmacology / Editorial Board, S.J. Enna (editor-in-chief) ... [Et al.], 2-16 (2016)
Proximal Ligation Assay (PLA) on Lung Tissue and Cultured Macrophages to Demonstrate Protein-protein Interaction
Mendez R, et al.
Bio-protocol, 7(21) (2017)



Global Trade Item Number

SKUGTIN
DUO82049-4L04061833591925
DUO82049-20L04061833591918