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Purification of His tagged protein


Mon Apr 21 13:30:53 EST 1997


Hi all,

I have been trying to purify a His tagged protein from Pichia -
unfortunately it is not secreted (but I am working on that) - however -
purification using Talon affinity resin or Ni agarose from crude extracts
yields loads of proteins? Has anyone succesfully purified by one step on
Talon resin - or should I try an initial purification step - size
exclusion or anion exchange? Being a bacteriologist (now a virologist) I'm
not aware if yeast have a large number of polyhistidine containing
proteins - can anyone help me with this?


Thanks

Ian Goodfellow




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