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Can a protein be eluted with EDTA from Ni-NTA affinity column...
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+ Elution
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+ Basic molecular biology techniques
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Posted by
Justin Siegel
Can a protein be eluted with EDTA from Ni-NTA affinity column...
I am doing purification of a 6xHis-tagged protein and it seems that the protein is not eluted with the traditional 500mM Imidazole in my elution buffer...mostly contaminating proteins are present. By coincidence (during column stripping and nickel recharging) I noticed a very big UV absorbance peak shortly after adding the stripping buffer. I collected and checked with SDS-PAGE. To my surprise there was my protein in high concentration. What would be the likely reason for this? Could it be the buffer composition and/or protein properties?
