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. Author manuscript; available in PMC: 2011 Sep 1.
Published in final edited form as: Hear Res. 2010 Jun 2;268(1-2):172–183. doi: 10.1016/j.heares.2010.05.019

Figure 1. Expression and purification of rOC90 and evaluation of glycosylation.

Figure 1

(A) Coomassie blue staining of purification fractions of rOC90. Lane 1. Molecular weight standard; Lane 2: Conditioned serum-free medium; Lane 3: Flow through; Lane 4: Final wash; Lane 5: Elution. (B) Western blot of transiently expressed rOC90 using monoclonal mouse anti-His antibody. Lane 1: intracellular rOC90; Lane 2: secreted rOC90. (C) PNGase F-treated rOC90. Lanes 1-3: Coomassie blue staining; Lanes 4-5: Western blot using polyclonal rabbit anti-mouse OC90 antibody. Lane 1: molecular weight standards; Lanes 2 and 4: rOC90; Lanes 3 and 5: PNGase F-treated rOC90. The PNGase F band is indicated by arrow. (D) GAG analysis of rOC90. Lanes 1-4: Coomassie blue staining; Lanes 5-8: Alcian blue staining. Lanes 1 and 5: Molecular weight standards; Lanes 2 and 6: 3 μg human fetuin A as positive control; Lanes 3 and 7: 3 μg rOC90; Lanes 4 and 8: 3 μg BSA as negative control.