Human fgf basic fgf2 bfgf duoset elisa

The levels of VEGF and bFGF present in conditioned media of MCF-7 and MDA-MB-231 cells were measured after 24 h of HA treatment by Human VEGF DuoSet ELISA Kit (R&D Systems, cat. no. DY293B-05, Minneapolis, MN, USA) and Human FGF basic/FGF2/bFGF DuoSet ELISA (R&D Systems, cat. no. DY233, Minneapolis, MN, USA) according to the supplier’s instructions.

The amounts of VEGF and FGF2 in CM were quantified using human VEGF DuoSet ELISA (R&D Systems, Minneapolis, MN, USA) and human FGF basic/FGF2/bFGF DuoSet ELISA (R&D Systems), following the manufacturer’s instructions. Optical density was measured using a microplate reader (450 nm, correction 540 nm, Infinite F50, Tecan, Mannedorf, Switzerland). The Proteome Profiler Human Angiogenesis Array Kit (R&D Systems) was used to analyze the expression of pro-angiogenic proteins in CM according to the manufacturer’s protocol. The pixel density of each spot was quantified using Image J software (National Institutes of Health), and the average signal was calculated for duplicate spots.