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. Author manuscript; available in PMC: 2012 Jan 25.
Published in final edited form as: Brain Res. 2010 Dec 16;1370:16–33. doi: 10.1016/j.brainres.2010.11.037

Figure 5. Estimation of molecular weights of recombinant NF95 and NF132 in Coomassie Blue-stained SDS gels.

Figure 5

The NF132 and NF95 genes were subcloned into the pET-11b (Novagen) expression vector and transfected into BL21(DE3)pLysS cells (Novagen). The cultures were harvested after incubation at 37°C overnight. Expression was induced by IPTG and controls were set without it. The cultures were allowed to grow for another 2 hours and lysed in homogenization buffer. The lysate proteins were separated on a 10% SDS-PAGE, stained with Coomassie Blue R-250 (0.1% in 50% methanol and 10% glacial acetic acid) and destained with 10% methanol and 7% acetic acid. A 95 kDa protein was expressed in NF95-transfected, IPTG-induced cells, while a 132 kDa protein was expressed in NF132-transfected, IPTG-induced cells (arrowheads).