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Documents
Quality Level
product line
Duolink®
application(s)
proximity ligation assay: suitable
fluorescence
λex 360 nm; λem 460 nm (Zeiss Filter set 49)
suitability
suitable for fluorescence
shipped in
wet ice
storage temp.
2-8°C
Related Categories
Application
Duolink® In Situ mounting medium with DAPI has been used to mount coverslips in proximity ligation assay (PLA) for nuclear staining.
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 Mounting Medium with DAPI is ideal for nuclear staining and preserving signals generated with the Duolink® In Situ Detection Reagents for fluorescence microscopy. See datasheet for more details.
Note: Counterstaining with Cy®2 is not recommended.
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
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 Mounting Medium with DAPI is ideal for nuclear staining and preserving signals generated with the Duolink® In Situ Detection Reagents for fluorescence microscopy. See datasheet for more details.
Note: Counterstaining with Cy®2 is not recommended.
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
Legal Information
Cy is a registered trademark of Cytiva
Duolink is a registered trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC
Signal Word
Warning
Hazard Statements
Precautionary Statements
RIDADR
NONH for all modes of transport
WGK Germany
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificate of Analysis
Certificate of Origin
S6K2-mediated regulation of TRBP as a determinant of miRNA expression in human primary lymphatic endothelial cells
Warner MJ, et al.
Nucleic Acids Research, 44(20), 9942-9955 (2016)
Intraneuronal Amylin Deposition, Peroxidative Membrane Injury and Increased IL-1β Synthesis in Brains of Alzheimer?s Disease Patients with Type-2 Diabetes and in Diabetic HIP Rats
Verma N, et al.
Journal of Alzheimer'S Disease, 53(1), 259-272 (2016)
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)
Takashi Miyamoto et al.
The Journal of biological chemistry, 291(4), 1719-1734 (2015-11-22)
Diverse lines of evidence suggest that amyloid-β (Aβ) peptides causally contribute to the pathogenesis of Alzheimer disease (AD), the most frequent neurodegenerative disorder. However, the mechanisms by which Aβ impairs neuronal functions remain to be fully elucidated. Previous studies showed...
Pengcheng Zhu et al.
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Cancer is a leading cause of death worldwide. Tumor cells exploit various signaling pathways to promote their growth and metastasis. To our knowledge, the role of angiopoietin-like 4 protein (ANGPTL4) in cancer remains undefined. Here, we found that elevated ANGPTL4...
Articles
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