An interesting puzzle - Pierce's "Gentle Ag/Ab elution buffer"? <-- SOLVED
noone at cancer.bham.ac.uk
Thu Apr 9 02:56:59 EST 1998
> Thanks very much to everyone who replied, to the group and by email.
> Many people suggested it could be magnesium. Harlow & Lane "Antibodies"
> book lists 3.5 M MgCl2 pH 7.2 as one of the alternatives to acid
> We ordered that stuff. It does not foam at all - so isn't detergent. It
> does form precipitate with phopshate, which is dissolved with EDTA but
> not EGTA, so it is Mg2+. By conductivity versus MgCl2 standards, it is
> 3.6 M solution. NaJO4 does not oxidize anything in this solution, so I
> don't think there is ethylene glycol present. Sooo, if anyone wants to
> make Pierce's magic solution, the recipie is:
> ~ 20 mM MES or PIPES,
> 3.6 M MgCl2,
> titrate pH to ~ 6.5 with NaOH
> Funny thing is that Pierce recommends to store this solution at 4C.
> Hehehe, like anything would grow in 3.5 M MgCl2!
> Or is it MgCl2 unstable? :-)
> 500 ml for $65 ... from Pierce
> or [when buying expensive MgCl2 from Sigma]
> 3.5 l for $104. Hmmm...
OK, but what's the brown precipitate I get from this buffer when I add
loading buffer (or any other DTT- or b-merc.-containing solution)?
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