Purification and Determination of some Characters of Dominant Isoform of GP96 Molecules from Mice Fibrosarcoma Cell Line (WEHI164)

Subject Areas : Journal of Comparative Pathobiology

عقیل Tabar Molla Hassan ¹ , زهیر Hassan ² , علی Mostafaie ³ , سعید Hesaraki ⁴ , علی Taravati ⁵

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Received: 2010-04-25 Accepted : 2010-10-07 Published : 2010-05-22

Keywords: affinity chromatography, ion exchange, purification,

Abstract :

GP96 protein is a heat shock protein (HSP) with an important role in cellular immunity. Tumorderived gp96 has entered clinical trials for cancer immunotherapy. For purification of dominantisoform of gp96 molecules from mice fibrosarcoma cell line, Cultured WEHI164 cell line werelysed and supernatants of cell extract were precipitated by Amoniume sulfate , In the next stage,purification was done through affinity and ione exchange chromatography. Molecular Weight ofpurified protein was estimated by SDS-PAGE and its nature was confirmed by western blot. Theresult of this study shows that, purified molecule has molecular weight between 66-68 kd whichindicates another different isoform of this molecule; a result confirmed by western blot.