Frequently Asked Questions

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Specificity

PLA probe anti-Rabbit reacts with whole molecule Rabbit IgG and the light chains of other Rabbit immunoglobulin′s.

Application

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.

To perform a complete Duolink in situ experiment you will need two primary antibodies (IHC or IF validated) that recognize two target epitopes. Additional reagents required include a pair of PLA probes, one PLUS and one MINUS , your choice of Detection Reagents. Optional reagents include Wash Buffers and Mounting Medium.

Analysis is carried out using standard immunofluorescence assay equipment. HRP/Novared is also available for bright field detection. Quick and reliable quantification can be performed using the Duolink Image Tool.

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) or immunohistochemistry (IHC) assay to determine the optimal fixation, blocking, and titer conditions.
Duolink in situ reagents are suitable for use on fixed cells, cyt-spin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). No minimum number of cells is required.

Features and Benefits

• Quicker than FRET
Visualize protein-protein interactions under endogenous expression
• Increased accuracy compared to co-IP
Single molecule detection capability due to >500X signal amplification
• Greater precision than current immuno assay techniques
Dual binding of primary antibodies enable greater specificity and low false positives

Legal Information

Duolink is a registered trademark of Olink AB

PLA is a registered trademark of Olink AB