Cell culture media, serum, and cell culture supplements were purchased from ATCC, USA, Merck Millipore, USA, LONZA, Switzerland and Thermo Fisher Scientific, USA. Antibodies against VE‐Cadherin (CD144) (ab33168 and STJ96234), VEGFR (ab9530), GAPDH (ab8245), OCT‐4 (ab19857 and STJ72238), KLF4 (ab72543) eNOS (ab76198), and FSTL3 (STJ112317) were purchased from Abcam, UK or St John's Laboratory, UK. Antibodies against vWF (SC‐8068) were purchased from Santa Cruz, USA. KDR (MAB3571), β‐actin (MAB8929), recombinant FSTL3 (AF1288‐F3), and Proteome Profiler Array Human Angiogenesis Array Kit (ARY007) were purchased from R&D, USA.
Proteome profiler array human angiogenesis array kit
Manufactured by R&D Systems
Sourced in United States
The Proteome Profiler Array Human Angiogenesis Array Kit is a multiplex assay used to detect the relative levels of 55 different human angiogenesis-related proteins in a single experiment. The kit includes a membrane-based assay, which allows for the simultaneous detection and quantification of multiple analytes from a single sample.
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Lab products found in correlation
3 protocols using proteome profiler array human angiogenesis array kit
1
Endothelial Cell Angiogenesis Assay
2
Paracrine Effects on Vascular Regeneration
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In order to examine the paracrine effects of Sk-34 and Sk-DN cells, key cytokines related to vascular regeneration were also analyzed by an antibody array kit (Proteome Profiler Array, Human angiogenesis array kit, ARY007, R & D Systems, Minneapolis, MN) a protein levels. Cell culture supernatants of Sk-34 and Sk-DN/29+ cells, prepared for use just prior to transplantation, were obtained after centrifugation, and 500 μl of the supernatants were prepared for the protein analysis. Culture medium containing 20% FCS was also prepared as the negative control. The relative protein expression levels of the selected angiogenesis-related cytokines, including Ang (Angiogenin), IGF binding protein (BP)-3, IL-8, MCP-1 (monocyte chemotactic protein-1), MMP-9, PlGF (placenta growth factor), and VEGF, were determined.
3
Angiogenesis Protein Array Analysis
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HUVECs were incubated with a 1:1 ratio of CM and ECM for 24 h and lysed in a buffer containing 1% NP-40, 20 mM Tris-HCl (pH 8.0), 137 mM NaCl, 10% glycerol, 2 mM EDTA and 1× protease inhibitor cocktail (Roche). 200 μg of the lysate was used per membrane of a Proteome Profiler Array, Human Angiogenesis Array Kit (R & D Systems). Three independent angiogenesis arrays with three independent protein sets from two lots of HUVECs were conducted. After visualizing the spots, the signals were measured with a Protein Array Analyzer plugin for ImageJ (https://imagej.nih.gov/ij/). Intensity files obtained from ImageJ were then uploaded to an in-house MATLAB code (MATLAB R2016a and Statistics Toolbox 11.0, The MathWorks, Inc., Natick, MA, USA, code available upon request). Details on normalizations methods are available in Supplementary Fig. 6.
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