Human angiogenesis growth factor magnetic bead panel kit

Analysis of various cytokines secreted by HAECs treated with VEGF (10 ng/mL) in absence or presence of aspalatone (50 μM and 100 μM) was analyzed by using a Human Angiogenesis/Growth Factor Magnetic bead panel kit from Millipore following manufacturer's instructions. Briefly, the culture media was collected, concentrated by lyophilization, and incubated with the labelled magnetic beads. After overnight incubation, the beads were counterstained with Streptavidin-Phycoerythrin and analyzed with a Luminex™ analyzer from Millipore. The results are expressed as pg/mL based on the standard curve generated with the standards.

Analysis of angiogenic cytokines secreted by HUVECs treated without or with VEGF (10 ng/mL) ± vialinin (5 μM) was performed by using a human angiogenesis/growth factor magnetic bead panel kit from Millipore (number HAGP1MAG-12 K) following the manufacturer's protocol. Briefly, after 24 h of treatment of HUVECS with VEGF ± vialinin A, the media were collected by centrifugation and filtered using a 0.2 μM syringe filter. The media were then frozen in −80°C and concentrated by a using a vacuum evaporator. Equal amounts of resuspended lyophilized media were then incubated with the labeled magnetic beads. After incubating overnight, the beads were counterstained with streptavidin-phycoerythrin and analyzed using a Luminex analyzer from Millipore. Data were analyzed using the xPONENT software, and the results are expressed as pg/mL.