Endothelial cells binding assay

Rot, A. (2003). In situ binding assay for studying chemokine interactions with endothelial cells. J Immunol Methods 273(1-2) 63-71. 

There are most likely a number of diverse molecular pathways by which NO can alter gene expression, but in the case of MCP-1 this pathway has been partially elucidated. In human endothelial cells inhibition of NO synthesis has been shown to activate proteins capable of binding to oligonucleotides containing the NF-kB binding site (Zeiher etal., 1995), suggesting a molecular link between an oxidant-sensitive transcriptional regulatory mechanism and NO synthesis. However, it cannot be excluded that NO also influences the activation of other transcription factors, such as AP-1, which has recently been characterized as an antioxidant-responsive factor (Meyer et al., 1993). Indeed, in in vitro gel-mobility assays the NO donor sodium nitroprusside, but no free NO, has been reported to S-nitrosylate the AP-1 moiety and inhibit its activity (Tabuchi et al., 1994). 

Caliper Technologies and Agilent have developed an annexin V assay in a microfluidic system, which allows flow cytometric analysis of apoptosis with a minimal number of cells [6, 7]. In this setup, the cells are moved by pressure-driven flow inside a network of microfluidic channels and are analyzed individually by two-channel fluorescence detection. As only a small number of cells (as few as 50-100) are consumed per assay, this setup is particularly suitable for working with cells of limited availability, for example, primary cells. The system has been applied to evaluate staurosporine-induced apoptosis and annexin V binding in human umbilical vein endothelial cells (HUVECs) and normal human dermal fibroblasts (NHDFs). The results are in good... 

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