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Latest discussion from ScientistSolutions.com

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  His-Tag Recombinant Protein purification

  Posted: January 31st, 2012, 8:30am EST

  Hello,
  I am purifying a six-His tagged 50kDa hnRNP K protein from E coli cells using IMAC with Cobalt TALON resin. I am trying to send the proteins for crystallography and it requires a good quality of sample. My problem is that smaller bands are visible on the gel and I don't know how to get rid of them.
  What might the problem and how can I fix it?
  Can using a whole range of gradients (50, 60, 70, 80, 90, 100, 110mM, etc.), or adding imidazole to the wash buffer improve...