TY  - JOUR
TI  - Proximal Ligation Assay (PLA) on Lung Tissue and Cultured  Macrophages to Demonstrate Protein-protein Interaction
AU  - Mendez, Roberto 
AU  - Banerjee, Santanu 
VL  - 7
IS  - 21
PY  - 2017
DA  - 2017/11/05
SP  - e2602
C1  - Bio-protocol 2017;7:e2602
DO  - 10.21769/BioProtoc.2602
UR  - https://doi.org/10.21769/BioProtoc.2602
AB  - In this protocol, we describe proximal ligation assay (PLA), an antibody-based detection method for protein-protein interaction. This method relies on specific binding of individual primary antibodies to the two putative interacting proteins. The primary antibodies need to have different hosts. The secondary antibodies against the two hosts have complementary oligonucleotide moieties attached to them. If the two antigens are in close proximity (presumably interacting with each other), the complementary oligonucleotides can anneal and fluorescent nucleotides can be incorporated in a single DNA polymerization step. Under a microscope, these reactions appear as punctate fluorescent spots, indicating successful PLA reaction and suggesting protein-protein interaction between the two antigens.
KW  - Proximal ligation assay
KW  - Protein-protein interaction
KW  - Toll-like receptors
KW  - Fluorescence microscopy
KW  - Bronchial epithelium
KW  - Macrophages
KW  - J774
KW  - Duolink
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False