We tried that trick, which works amazingly well in insect cells, in
mammalian media, and it fails. It will depend on the exact media, obviously.
Engin
On 10/8/09 1:50 PM, Matthew Franklin wrote:
The trick we used at Genentech (which I'm still using) was for
secreted insect cell proteins, but it should work for you as well. Add
1 mM NiCl2 and 10 mM CaCl2 to your conditioned media, and adjust the
pH to 7.2 - 7.5. For insect cell conditioned media, this produces a
fairly heavy precipitate which appears to contain the nickel-chelating
factor. Most of the time (but not always!) your protein of interest
will not be precipitated by this step, and you can spin out the
precipitate then load the supernatant on to your nickel column with no
further processing.
Hope that helps,
Matt
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Engin Özkan
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