Roger Murphy <roger.murphy at ludwig.edu.au> wrote:
:I seem to remember some discussion of "gentle elution buffer" for
:eluting antibodies from protein A some time ago - if my memory serves me
:correctly, it was basically magnesium chloride solution. Does anyone
:remember the recipe for this buffer?
:
Yes, I remember because I posted it :-)
3.5 - 3.6 M MgCl2, pH 6.5 as measured by a regular calomel
electrode (naturally, at this salt conc. the true pH should be
considerably different). The main disadvantage is you can't
really measure protein conveniently in the presence of
3.5 M Mg2+ - Bradford loses sensitivity, BCA precipitates
Not sure what they buffer it with or even if they do, but
anything will work and I am told by my wife that in her hands
pH 7.0 appears to be as effective. In my hands, the elution works
a lot better at rt. In the cold, fractions tend to "tail" a lot
more. Make sure to leave plenty of free space in the bag
when you dialyze.
- Dima
P.S. Isn't it funny that Pierce recommends storing this solution
in the cold? Like anything can grow in 3.5 M MgCl2!
