The Duolink® using PLA® Technology Product line has not been tested with whole mount samples.
DUO92012
Duolink® In Situ Detection Reagents Brightfield
Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
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30 REACTIONS
€ 540,00
100 REACTIONS
€ 807,00
About This Item
UNSPSC Code:
12352200
NACRES:
NA.32
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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
1 of 4
This Item | DUO92007 | DUO82049 | DUO92010 |
|---|---|---|---|
| suitability suitable for brightfield | suitability suitable for fluorescence | suitability suitable for fluorescence | suitability suitable for brightfield, suitable for fluorescence |
| technique(s) proximity ligation assay: suitable | technique(s) proximity ligation assay: suitable | technique(s) proximity ligation assay: suitable | technique(s) immunofluorescence: suitable, proximity ligation assay: suitable |
| storage temp. −20°C | storage temp. −20°C | storage temp. 20-25°C | storage temp. −20°C |
| product line Duolink® | product line Duolink® | product line Duolink® | product line Duolink® |
| packaging pkg of Box A (-20°C) | packaging - | packaging - | packaging - |
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
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Acute Tox. 3 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3 - Eye Dam. 1 - Flam. Liq. 2 - Muta. 2 - Resp. Sens. 1 - Skin Irrit. 2 - Skin Sens. 1
Storage Class Code
3 - Flammable liquids
Flash Point(F)
42.8 °F
Flash Point(C)
6 °C
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Agata Zieba et al.
Clinical chemistry, 56(1), 99-110 (2009-11-21)
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
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Articles
Erfahren Sie, wie die Proximity Ligation Assay-Technologie funktioniert und wie durch das Kontrollkit für die Protein-Protein-Interaktion der In-situ-Nachweis der EGF-induzierten EGFR-HER2-Dimerisierung bestätigt werden kann.
Support information including tips and tricks, frequently asked questions, and basic troubleshooting.
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.
Things to consider for preparation, setup and execution of the Duolink® assay protocol
Protocols
Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.
Related Content
Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay
Instructions
Application Note
Using Duolink in Multiwell Plates
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Is it possible to utilize Duolink® with PLA® Technology for whole mount or thick tissue samples?
1 answer-
Helpful?
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I received 2 DuoLink kits, DUO92012A-100RXN and DUO92012B- 100RXN, each containing the kit component DUO82057 lot SLCN4945. One of the bottles of DUO82057 lot SLCN4945 is brown, while the other is black. Please provide troubleshooting assistance for this issue?
1 answer-
The supplier has confirmed that the reagent is still suitable for use and that darkening over time is expected, with no impact on the quality or intensity of staining. The variation in coloration between the same lot is likely due to differences in the amount of time each reagent has been opened and exposed, leading to oxidation.
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What are the suitable stopping points in the Duolink PLA protocol that will not have a negative impact on the results?
1 answer-
The Duolink® PLA protocol offers several stopping points, allowing samples to be left at various stages. For instance, after fixation, samples can be left in 1x PBS at 4 °C for several days. Additionally, blocking and primary antibody incubations can be run overnight. Once the procedure is finished, slides can be stored at different temperatures for varying durations based on the detection method used. It is important to note that the ligation and amplification conditions have been optimized and should not be altered, unless for specific applications where extra caution is necessary.
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