Duolink™ In Situ Detection Reagents Green
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.
Green fluorescence detection reagents are often used with FITC filter.
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.[3][1][2][4]
• 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
• 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.
See datasheet for more information.
Not included in Detection kit:
Primary
antibodies, PLA probes, wash buffers, mounting medium
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.
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.
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
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Cy is a registered trademark of Cytiva
Duolink is a trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC
