Abstract
Proteins differ widely in their pattern of expression depending on organism, tissue, and regulation in response to changing conditions. In the mammalian vasculature, the endothelium responds to vascular endothelial growth factors (VEGFs) via membrane-bound receptor tyrosine kinases (VEGFRs) to modulate many aspects of vascular physiology including vasculogenesis, angiogenesis, and blood pressure. Studies on VEGFR biology are thus dependent on detecting expression levels in different cell types and evaluating how changes in protein levels correlate with changing conditions including circulating VEGF levels. Here, we present a robust immunoblot-based protocol for detecting and quantifying VEGFRs in human endothelial cells. Using internal and external standards, we can rapidly evaluate receptor copy number and assess how this is altered in response to the cellular environment.
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Acknowledgements
This work was supported by a Heart Research UK PhD studentship (G.W.F.) and British Heart Foundation project grants (S.P. and S.B.W.).
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Fearnley, G.W., Wheatcroft, S.B., Ponnambalam, S. (2015). Detection and Quantification of Vascular Endothelial Growth Factor Receptor Tyrosine Kinases in Primary Human Endothelial Cells. In: Fiedler, L. (eds) VEGF Signaling. Methods in Molecular Biology, vol 1332. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2917-7_4
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DOI: https://doi.org/10.1007/978-1-4939-2917-7_4
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2916-0
Online ISBN: 978-1-4939-2917-7
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