An interesting puzzle - Pierce's "Gentle Ag/Ab elution buffer"? <-- SOLVED

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Wed Apr 8 12:20:04 EST 1998


In article <6g499h$1ek_002 at doit.wisc.edu>, 
klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu (Dima Klenchin) wrote:
:I am trying to understand what that could be...
:
:        What is known: 
:
:efficiently disrupts antigen/antibody interactions, pH is 6.5, is incompatible 
:with phosphate, is not chaotropic agent, is dialyzable out (the latter told by 
:Pierce tech support, so is not nesessarily correct :-)) .
:
:I am pulling my hairs out trying to understand what that could be... My best 
:guess is some mild ionic detergent like CHAPSO and the likes but 
:incompatibility with phosphate stumbles me...

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 
elution. 

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...

        Dima








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