Cobalt turning orange ??

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Cobalt turning orange ??

Post by mgn67 » Sun Jan 22, 2012 11:32 am


I've been trying to purify a his-tagged protein, using HiTrap IMAC column loaded with CoCl2. And everytime, my protein vanishes : it's there in the soluble extract that i load on the column, but not anymore after the column, neither in the flow through or the elution...
I've been then trying the old batch method... In parallel, with Talon resin (cobalt) and Ni-NTA resin (nickel). What was weird, was a change of colour of the Talon resin : it turned light orange with my sample...
Final result, my protein is pure from Ni-NTA resin but no results from Talon resin...
Has someone already observed something similar, or has some explanation on this change of colour ?

I doubt it's due to the buffer (500mM NaCl, 20mM Tris 7.5, 1mM TCEP, 5% glycerol, 0.01% Tween-20) cause i purified an other protein with the same buffer, and same HiTrap IMAC column with cobalt, and the purification was perfect... Could it be due to the protein itself ?



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