How can I make an elution to extract a protein on an IgG bead without washing off the protein?

11 July 2018 1 2K Report

I used GST-3C protease and Glutathione Sepharose 4B beads that were washed 3X with 3C buffer. Then everything was filtered through a SPIN-X centrifuge tube. When I ran the Coomassie Blue gel, I did not get any protein bands. When I keep the protein on the IgG bead, I get good levels of expression in both the Western Blot and the Coomassie gel. Any suggestions?

Luis PARMENIO Suescún Bolívar

you should test a elution with salt (NaCl) gradient.

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