Download - Proximity Ligation And RCA
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Direct observation of individual
endogenous protein complexes
in situ by proximity ligation
SÖderberg et al., Nature Methods, 2006, 3, 995.
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Science, 1994, 265, 2085.
Rolling Circle Replication
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Nucleic Acids Research, 1998, 26, 5073.
Rolling Circle Replication
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Nature Biotechnology, 2002, 20, 448.
Proximity ligation
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Nature Methods, 2006, 3, 995.
Rolling Circle Replication/Proximity ligation
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Nature Methods, 2006, 3, 995.
U2OS Skov-3
CHO-K1 HFB
TIME CELLS
Mitotic Cells
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Nature Methods, 2006, 3, 995.
Rat1 TGR-1 fibroblast
Rat1 TGR-1 fibroblastc-myc knockout
C-Myc / EGFR proximity probes Her-2 / EGFR proximity probes
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Nature Methods, 2006, 3, 995.
U2OS stained with 2 different hybridization probes
SHSY5Y cells
Identification of interaction of c-Myc/Max with RNA Pol II
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Nature Methods, 2006, 3, 995.
Normal colon tissue
Normal colon tissue(Enzyme based immunostaining)
Human tonsil tissues
Burkitt Lymphoma sample
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Nature Methods, 2006, 3, 995.
U-937 cells transformed with v-myc
U-937 cells transformed with v-myc + PMA + IFN-
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Nature Methods, 2006, 3, 995.
4-OH-Tamoxifen: Antagonist to ER
Small molecule inhibitors of c-Myc/Max interaction
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Conclusions
-P-LISA can detect protein-protein interaction at the single molecule level.-RCA increases the number of fluorophores/detected interaction.-Multi protein complexes can be studied.-Doesn’t need expression of fusion proteins.-Disruption of protein-protein interaction can be studied.
-P-LISA can be performed only on fixed cells-Real time tracking cannot be possible-Selection of antibodies for each pair
-Validate results from Yeast Two Hybrid or other assays-Can be used as molecular ruler