All Photos(4)
Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
UNSPSC Code:
12352200
NACRES:
NA.32
Recommended Products
product line
Duolink®
Quality Level
technique(s)
proximity ligation assay: suitable
fluorescence
λex 495 nm; λem 527 nm (green) (FITC (Cyanine 2), Zeiss Filter set 38)
suitability
suitable for fluorescence
shipped in
dry ice
storage temp.
−20°C
Related Categories
General description
Duolink® In Situ Detection Reagents Green contains all the necessary Duolink In situ reagents to perform the amplification and detection of bound PLA® probes. The detection probes contain a fluorophore (lex = 495 nm and lem = 527 nm), which may be visualized using the same filter as Cy®2 or FITC. Experiments conducted using Duolink In situ reagents can detect and visualize protein interactions, protein expression levels and post translational modifications at the single molecule level in fixed cells and tissue samples.
Specificity
Green fluorescence detection reagents are often used with FITC filter.
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.
Duolink® In Situ Detection Reagents has been used in the proximity ligation assay of:
- vasopressin and gonadotropin-releasing hormone (GnRH) from frozen rat brain sections
- human embryonic kidney 293 cells (HEK)
- pituitary tissues
Features and Benefits
- No overexpression or genetic manipulation reNo 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
Components
- 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
- 1x Ligase (1 unit/μL)
- 1x Polymerase (10 units/μL)
- 5x Amplification Green - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase. It also contains oligonucleotide probes labeled with a fluorophore that hybridize to the RCA product.
Not included in Detection kit:
Primary antibodies, PLA probes, wash buffers, mounting medium
Preparation Note
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.
Storage and Stability
Store the 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.
Other Notes
Follow the Duolink® In Situ Fluorescence Protocol to use 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.
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects
View full Duolink® product list
Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects
View full Duolink® product list
Legal Information
Cy is a registered trademark of Cytiva
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
Resp. Sens. 1
WGK
WGK 2
Regulatory Information
常规特殊物品
含少量动物源组分生物产品
Certificates of Analysis (COA)
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Articles
Things to consider for preparation, setup and execution of the Duolink® assay protocol
Support information including tips and tricks, frequently asked questions, and basic troubleshooting.
Protocols
This protocol describes how to perform immunofluorescent detection of proteins in cells and tissue.
This page details the Duolink® In Situ Short Protocol for fluorescence detection
Related Content
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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