In <199407131917.MAA06270 at net.bio.net> ERNESTO at VM.CC.PURDUE.EDU (Martin Tiznado) writes:
>I am working in Pectin Methyl Esterase and I had observed that when I use low
>pH (2.5) to elute the protein from an affinity column, I always lost one of the
> proteins. This phenomenon does not happen with another procedures. Any ideas a
>bout. Hint: Those proteins can be glycosilated.
What kind of affinity column do you use? A couple of times I couldn't
elute proteins from immunoaffine columns with Gly or acetic acid, instead I
used high pH buffers. Do you loose protein or activity? Do you
collect eluates right into Tris in order to neutralize acid? Some enzymes are
pretty sensitive to low pH values. How about proteases in your extracts?
AR
