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Characterization of GPCRs in extracellular vesicle (EV).

Methods Cell Biol.. 2017; 
Medapati MR, Singh A, Korupally RR, Henderson D, Klonisch T, Manda SV, Chelikani P.
Products/Services Used Details Operation
... 6. Antibodies used: Anti-CD63 mouse monoclonal (Abcam, #ab59479), Anti-Flag-M2 monoclonal(Sigma, #F1804), Anti-Avi mouse monoclonal (Genscript, #A01738), Normal mouse IgG (Upstate cell signaling solutions, #12-371), Goat antimouse secondary (10 nm gold ...

摘要

Extracellular vesicle (EV) are tiny membranous vesicles usually <500nm in size that recently emerged as a new paradigm in human intercellular signaling. EVs have shown a promising role in development of diagnostic markers in many pathophysiological disorders. The presence of chemosensory and therapeutically relevant G protein-coupled receptors (GPCRs) on EV membranes is poorly characterized. Here, we compare different methods including ultracentrifugation and polymer-charge-based separation to isolate EVs from cell culture media and human saliva. The presence of bitter taste GPCRs (T2R4 and T2R38) and a class A GPCR angiotensin II type 1 receptor on these EVs was characterized by qPCR, ELISA, and immunotransmis... More

关键词

Angiotensin II type 1 receptor; Bitter taste receptor; Electron microscopy; Exosomes; Extracellular vesicle; G protein-coupled receptor; Polyethylene glycol; Protamine chloride