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An Ultra‐High Fluorescence Enhancement and High Throughput Assay for Revealing Expression and Internalization of Chemokine Receptor CXCR4
Author(s) -
He Hua,
Wang Xiaojuan,
Cheng Tiantian,
Xia Yongqing,
Lao Jun,
Ge Baosheng,
Ren Hao,
Khan Naseer Ullah,
Huang Fang
Publication year - 2016
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201600245
Subject(s) - internalization , chemokine receptor , alexa fluor , cancer cell , cxcr4 , chemistry , chemokine , biophysics , microbiology and biotechnology , fluorescence , materials science , receptor , cancer , biology , biochemistry , physics , quantum mechanics , genetics
Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high‐throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip‐coating poly(styrene‐co‐ N ‐isopropylacrylamide) spheres on the glass substrate. The self‐ assembled spheres form three‐dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF‐1α activation in various cancer cells, even for those with extremely low expression level.