PMID- 28117799
OWN - NLM
STAT- MEDLINE
DCOM- 20170927
LR  - 20190116
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 119
DP  - 2017 Jan 16
TI  - Visualization of Protein-protein Interaction in Nuclear and Cytoplasmic Fractions 
      by Co-immunoprecipitation and In Situ Proximity Ligation Assay.
LID - 10.3791/55218 [doi]
LID - 55218
AB  - Protein-protein interactions are involved in thousands of cellular processes and 
      occur in distinct spatial context. Traditionally, co-immunoprecipitation is a 
      popular technique to detect protein-protein interactions. Subsequent Western blot 
      analysis is the most common method to visualize co-immunoprecipitated proteins. 
      Recently, the proximity ligation assay has become a powerful tool to visualize 
      protein-protein interactions in situ and provides the possibility to quantify 
      protein-protein interactions by this method. Similar to conventional 
      immunocytochemistry, the proximity ligation assay technique is also based on the 
      accessibility of primary antibodies to the antigens, but in contrast, proximity 
      ligation assay detects protein-protein interactions with a unique technique 
      involving rolling-circle PCR, while conventional immunocytochemistry only shows 
      co-localization of proteins. Nuclear factor 90 (NF90) and RNA-binding motif 
      protein 3 (RBM3) have been previously demonstrated as interacting partners. They 
      are predominantly localized in the nucleus, but also migrate into the cytoplasm 
      and regulate signaling pathways in the cytoplasmic compartment. Here, we compared 
      NF90-RBM3 interaction in both the nucleus and the cytoplasm by 
      co-immunoprecipitation and proximity ligation assay. In addition, we discussed 
      the advantages and limitations of these two techniques in visualizing 
      protein-protein interactions in respect to spatial distribution and the 
      properties of protein-protein interactions.
FAU - Zhu, Xinzhou
AU  - Zhu X
AD  - University Children's Hospital Basel (UKBB), University of Basel; 
      xinzhou.zhu@ukbb.ch.
FAU - Zelmer, Andrea
AU  - Zelmer A
AD  - University Children's Hospital Basel (UKBB), University of Basel.
FAU - Wellmann, Sven
AU  - Wellmann S
AD  - University Children's Hospital Basel (UKBB), University of Basel; Department of 
      Clinical Research, University of Basel.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20170116
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Antibodies)
RN  - 0 (ILF3 protein, human)
RN  - 0 (Nuclear Factor 90 Proteins)
RN  - 0 (Proteins)
RN  - 0 (RBM3 protein, human)
RN  - 0 (RNA-Binding Proteins)
SB  - IM
MH  - Antibodies
MH  - Blotting, Western
MH  - Humans
MH  - *Immunoprecipitation
MH  - Nuclear Factor 90 Proteins/chemistry
MH  - Protein Interaction Mapping/*methods
MH  - Proteins/*chemistry
MH  - RNA-Binding Proteins/chemistry
MH  - Signal Transduction
PMC - PMC5352272
EDAT- 2017/01/25 06:00
MHDA- 2017/09/28 06:00
PMCR- 2019/01/16
CRDT- 2017/01/25 06:00
PHST- 2017/01/25 06:00 [entrez]
PHST- 2017/01/25 06:00 [pubmed]
PHST- 2017/09/28 06:00 [medline]
PHST- 2019/01/16 00:00 [pmc-release]
AID - 55218 [pii]
AID - 10.3791/55218 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Jan 16;(119):55218. doi: 10.3791/55218.