Try adding 6 mg mercaptoethylamine hydrochloride to antibody in 100mM
phosphate pH ~7, incubate 37 deg about 2h. Desalt into a buffer containing
5mM EDTA, which I understand prevents metal catalysed oxidation back to the
disulfides (I generally feel safer if I first vacuum degas the buffer a few
minutes). This is from memory of a procedure in a Pierce brochure which came
with the MEA, but should be pretty close.
John
Nigel Osborn wrote:
> I want to fragment an antibody at the disulfide bond between the two heavy
> chains leaving all the other disulfides intact. This gives a half IgG
> (univalent) with molecular weight around 75kD.
>> Does anyone know a method for doing this?
>> Thanks.
>> Nigel.
