I have tried to purify alpha synuclein from DEAE-sepharose column. In that cell pellet are lysed again osmotic buffer. And then pellet collected after centrifugation, was resuspended in cold water. After further centrifugation, lysate was collected which is loaded into DEAE-sepharose matrix and eluted using 0.5 M Nacl.
In this particular protocol, i found non specific bands are coming after elution. Please give me suggestion regarding this.
Non-specific bands mean bands which are coming at the location other than near 15-20 kDa in 15% SDS-PAGE.
You are using an ion exchange system, which is not specific for similarly charged proteins. Run a nacl gradient to separate adsorbed proteins
Thank you for valuable suggestion. I will definitely try this method.
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