Duolink™ In Situ PLA® Probe Anti-Human MINUS
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.
This product can be applied to both the Duolink™ In Situ Fluorescence Protocol and the Duolink™ In Situ Brightfield Protocol depending on the detection reagents used.
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. HRP is also available for brightfield detection.
Specificity
PLA probe anti-Human reacts with whole molecule
human IgG and the light chains of other human immunoglobulins. The PLA Probe Anti-Human Minus has minimal cross-reactivity with bovine, chicken, goat, guinea pig, Syrian hamster, horse, mouse, rabbit, rat, and sheep serum proteins. A PLUS probe of
a different species must be used simultaneously with this product. See our Product Selection Guide for more information.
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™ PLA 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.
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• 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
This product is comprised of the following:
• 5x PLA Probe Anti-Human MINUS - Donkey anti-human secondary antibody conjugated to oligonucleotide MINUS
• 1x Blocking Solution - Reagent for blocking of the sample
• 1x Antibody
Diluent - For dilution of PLA probes and primary antibodies
See datasheet for more information.
Duolink is a trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC
