Fearnley, GW, Wheatcroft, SB and Ponnambalam, S orcid.org/0000-0002-4452-7619 (2015) Detection and Quantification of Vascular Endothelial Growth Factor Receptor Tyrosine Kinases in Primary Human Endothelial Cells. Methods in Molecular Biology, 1332. pp. 49-65. ISSN 1064-3745
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Keywords: | Human umbilical vein endothelial cells (HUVECs); VEGFR1; VEGFR2; VEGFR3; Quantification; Immunoblot; SDS-PAGE |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 30 Sep 2016 14:26 |
Last Modified: | 04 Nov 2016 07:36 |
Published Version: | http://dx.doi.org/10.1007/978-1-4939-2917-7_4 |
Status: | Published |
Publisher: | Springer |
Identification Number: | 10.1007/978-1-4939-2917-7_4 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:96535 |