The in situ proximity ligation assay is a powerful technology capable of detecting single protein events such as protein protein interactions (e.g. protein dimerization) and modifications (e.g. protein phosphorylation) in tissue and cell samples prepared for microscopy. Each detected signal is visualized as an individual fluorescent dot, these signals can be quantified (counted) and assigned to a specific subcellular location based on microscopy images. This revolutionary technique enables unprecedented specificity and sensitivity of protein detection and quantification for high performance immunofluorescence and immunohistochemistry applications. Leveraging on Abnova's manufacturing capability and growing collection of 50,000+ antibodies, hundreds of target specific and validated assays are being developed in cells and tissue samples.
Principle of Proximity Ligation Assay
1.
Incubate with target primary antibodies from two different species
2.
Add probes
3.
Hybridize connector oligos
4.
Ligation to form a complete DNA circle
5.
Rolling circle amplification
6.
Add fluorescent probes to reveal protein protein interaction
Huh7 cells were stained with anti-FLT1
rabbit purified polyclonal antibody and
anti-CTNNB1 mouse monoclonal
antibody. Each red dot represents the
detection of protein-protein interaction
complex.
HeLa cells were stained with anti-
TRAF2 rabbit purified polyclonal
antibody and anti-ACTG1 mouse
monoclonal antibody. Each red dot
represents the detection of protein
protein interaction complex.
HeLa cells were stained with anti-SNCA
rabbit purified polyclonal antibody and
anti-BAX mouse monoclonal antibody.
Each red dot represents the detection of
protein-protein interaction complex.
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