How is the in situ proximity ligation assay used?
In situ proximity ligation assay (PLA) is a method to identify physical closeness of proteins, where a signal will only be produced if the two proteins are closer than 40 nm, in tissue section or cell cultures. Modifications of the PLA method can also be used to increase specificity or sensitivity of standard immunohistochemistry protocols. Animals
How does duolink proximity ligation assay ( PLA ) work?
Duolink ® Proximity Ligation Assay (PLA) is a powerful tool that allows in situ detection of endogenous proteins, protein modifications, and protein interactions with high specificity and sensitivity. Protein targets can be readily detected and localized with single molecule resolution in unmodified cells and tissues.
How are PLA probes ligated in proximity ligation?
Briefly, when the PLA probes are in a sufficiently close distance, the oligos connect with the PLA probes and are ligated. There are two PLA probes: PLUS and MINUS. The PLA probe PLUS binds to the antibody recognizing one monomer, while MINUS binds to antibody recognizing second protomer.
How does immobilization of a protein help in ligation?
Immobilization of the protein on a solid support can serve to purify and concentrate the samples, especially those that contain constituents that may inhibit ligation or amplification, or where the concentration of analyte is extremely low.
How is duolink PLA used in protein detection?
Rely on Duolink ® PLA to amplify protein signals by 1000x, allowing you to visually detect proteins and their interactions, all with a straightforward protocol. Uncover the opportunities in your research with amplified performance and the benefits of Duolink ® PLA technology:
What can duolink probemaker kits be used for?
Duolink ® Probemaker Kits enable the conjugation of a PLA oligo (PLUS or MINUS) to any antibody. Duolink ® PLA Multicolor Kits allow for multiplex applications. Duolink ® Fluorescent Kits enable use with four different fluorophores: green, orange, red, and far red.
Where do you store duolink fluorescent detection reagent?
Duolink ® Fluorescent Detection Reagent (choose from green, orange, red, and far red). Each kit includes: NOTE: Store all components at -20 °C. Duolink ® Mounting Media with DAPI (for use on slides, store at 2-8 °C) OR Nuclear Stain and Anti-fade (for use in microplates, store at -20 °C)