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A rapid method for detection of cell adhesion molecules (CAMs) on human umbilical vein endothelial cells (HUVECs)
Authors:Saroj Kumar  Sarvesh Kumar  Pradip Nahar
Affiliation:a Institute of Genomics and Integrative Biology (CSIR), Mall Road, Delhi 110007, India
b University of Iowa, Iowa City, IA, USA
Abstract:Detection of cell surface proteins is widely used as molecular markers for initiation, progression and severity of many diseases. In particular, detection of cell adhesion molecules (CAMs) on endothelial cells is important as it indicates the extent of inflammation associated with several diseases including arthritis, asthma, tumor metastasis, etc. Here, we report, a rapid method for detection of CAMs on endothelial cells by covalently immobilizing TNF-α induced cells on a photoactivated polystyrene microtiter plate at 50 °C in 45 min followed by performing enzyme-linked immunosorbent assay (ELISA) technique at elevated temperature. Our method reduced the time of cell-ELISA to 3 h with results akin to conventional cell-ELISA carried out in 38 h. The method thus described herein could be potentially useful in clinical and research laboratories for rapid detection of cell surface proteins including CAMs on intact cell samples.
Keywords:ICAM-1, intercellular adhesion molecule-1   VCAM-1, vascular cell adhesion molecule-1   BSA, bovine serum albumin   FNAB, 1-fluoro-2-nitro-4-azidobenzene   HUVECs, human umbilical vein endothelial cells   HELISA, heat-mediated enzyme-linked immunosorbent assay   cell-HELISA, heat-mediated cell-ELISA   EC, endothelial cell   OPD, o-phenylenediamine dihydrochloride
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