Fig 2: Western Blot Analysis of sVEGFR1 in hUTC Conditioned MediumThe eluates of hUTC CM and control medium from VEGF pull-down assay were subject to SDS-PAGF. Briefly, 40 uL Tris-eluate solution mixed with sample buffer and 25 ng recombinant human soluble VEGFR1 were run on an SDS-PAGE gel, and the gel-resolved proteins were transferred onto a PVDF membrane. The membrane was blocked with 5% (w/v) BSA, and subsequently it was probed with a biotinylated anti-VEGFR1 antibody targeting the extracellular region of VEGFR1. After washing, the membrane was incubated with streptavidin-HRP, and immunoreactive bands were visualized with enhanced chemiluminescence (ECL). CON, hUTC control medium sample; NEG, negative control (control medium); REC, recombinant human sVEGFR1.