I have used heparin column several times to purify DNA binding protein and it seems that the purification quality has now reduced. Initially only protein of interest used to bind and elute from the column and now it is getting eluted with other non-specific proteins.
I have looked on the GE website but they have not provided any information.
Does anyone have regenerated the column? Kindly provide some suggestions.
How well do you isolate your POI before heparin binding and how monomeric is your POI entering the heparin column? Sometimes that can play a role in what comes out during elution. Several column volumes of 1M NaCl should be enough clear any soluble protein. However, we use 0.1M NaOH to "clean" our HiTrap Heparin columns about once a month depending on usage.
https://www.sigmaaldrich.com/technical-documents/protocols/biology/affinity-chromatography/separating-dna-binding-proteins-with-heparin.html
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