DUO92012
Duolink® In Situ Detection Reagents Brightfield
Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
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About This Item
NACRES:
NA.32
UNSPSC Code:
12352200
product line
Duolink®
packaging
pkg of Box A (-20°C), pkg of Box B (2-8°C)
technique(s)
proximity ligation assay: suitable
suitability
suitable for brightfield
storage temp.
−20°C
Application
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.
Follow the Duolink® In Situ Brightfield Protocol to use this product.
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 HRP for brightfield detection.
Follow the Duolink® In Situ Brightfield Protocol to use this product.
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 HRP for brightfield detection.
Specificity
Brightfield detection reagents are used for samples with significant autofluorescence.
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® 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.
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects
View full Duolink® product list
Brightfield detection reagents are used for samples with significant autofluorescence.
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® 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.
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects
View full Duolink® product list
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
Other Notes
Storage/Stability
Store Box A and all of its components at -20°C. The enzymes should be kept cold (-20°C) at all times; Use a freezing block when removing them from the freezer.
Store Box B and all of its components at 2-8°C.
Store Box A and all of its components at -20°C. The enzymes should be kept cold (-20°C) at all times; Use a freezing block when removing them from the freezer.
Store Box B and all of its components at 2-8°C.
This product is comprised of the following:
Box A:
Box B:
Not included in Detection kit:
Primary antibodies, PLA probes, wash buffers, mounting medium
Box A:
- 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
- 1x Ligase (1 unit/μL)
- 5x Amplification - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase
- 5x Detection Brightfield - Contains oligonucleotide probes labeled with horseradish peroxidase (HRP) that hybridize to the RCA product
- 1x Polymerase (10 units/μL)
Box B:
- 1x Hydrogen Peroxide - Contains 0.3% hydrogen peroxide
- Substrate Reagents A-D - Contains all substrate components needed for HRP enzymatic reaction
- 1x Nuclear Stain - Contains Mayer′s hematoxylin solution for cell nuclei staining
Not included in Detection kit:
Primary antibodies, PLA probes, wash buffers, mounting medium
Legal Information
Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany
signalword
Danger
Storage Class
3 - Flammable liquids
flash_point_f
42.8 °F
flash_point_c
6 °C
wgk
WGK 3
Hazard Classifications
Acute Tox. 3 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 2 - ED ENV 1 - Eye Dam. 1 - Flam. Liq. 2 - Muta. 2 - Resp. Sens. 1 - Skin Irrit. 2 - Skin Sens. 1
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Protocols
使用明场显微镜和Duolink® PLA试剂用于检测和定量组织和细胞样品中的个体蛋白质、蛋白质修饰和蛋白质相互作用。
Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.
Articles
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。
了解邻位连接技术的工作原理以及蛋白质互作对照试剂盒如何确定EGF诱导的EGFR-HER2二聚化的原位检测。
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。
Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay
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The in situ proximity ligation assay (PLA) allows a protein or protein complex to be represented as an amplifiable DNA molecule. Recognition is mediated by proximity probes consisting of antibodies coupled with oligonucleotides. Upon dual binding of the proximity probes
Global Trade Item Number
| SKU | GTIN |
|---|---|
| DUO92012-100RXN | 04061833592137 |
| DUO92012-30RXN | 04061833592144 |