Fig. 2
Effect of culture conditions on the expression of VEGF165 in the pellet (insoluble) and the supernatant (soluble) of centrifuged crude lysates. a Insoluble VEGF165 expression at different induction temperature, different IPTG concentration and different medium. Lanes 1–3: insoluble VEGF165 expression induced with 0.4 mM IPTG in 2YTA medium at the temperature 25, 30 and 37 °C respectively. Lane 4 insoluble VEGF165 expression induced with 1 mM IPTG at 37 °C in 2YTA medium. Lanes 5–7 insoluble VEGF165 expressed in LBA medium and induced with 0.4 mM IPTG at 25, 30 and 37 °C respectively. Lane M protein marker (GE Healthcare UK limited). Lane 8 empty pET 21a vector lysate (control). b SDS-PAGE (top) and Western blot (bottom) analysis of the soluble VEGF165 expression at different induction temperature, different IPTG concentration and different medium. Lane 1–3 soluble VEGF165 expression induced with 0.4 mM IPTG in 2YTA medium at the temperature 25, 30 and 37 °C respectively. Lane 4 soluble VEGF165 expression induced with 1 mM IPTG at 37 °C in 2YTA medium. Lanes 5–6 soluble VEGF165 expressed in LBA medium and induced with 0.4 mM IPTG at 25 and 37 °C respectively