Re: nonionic detergents
Joseph Fernandez (fernaj@rockvax.rockefeller.edu)
Mon, 4 May 1998 09:39:06 -0400 (EDT)
Vernon,
Just got back into the lab and didn't notice your question till
this mourning. I am not sure I know of too many techniques for analysis of
nonionic detergents, but I do know what effect they have on RP-HPLC,
particularly regarding peptide mixtures.
1) Nonionic detergetns can have uv absorbing artifacts due to heterogeneity
but there are several that can be used that are uv transparrent. For
example, the reduced (hydrogenated) form of Triton X-100 is basically uv
transparrent as opposed to the nonhydrogentaed form Triton X-100.
2) Other nonionics such as Genepol X-180, Tween-20, Tween-80,
octylglucopyranoside are also useful to RP-HPLC, as well as Zwitterionic
detergents.
3) When analyzing peptides on RP-HPLC using nonionics mentioned above,
there is no change in retention time, peak resolution, or peptide recovery.
Here is a list of papers utilizing nonionics for RP-HPLC
Fernandez et al, Anal Biochem, 1992, 201, 255-264.
Fernandez et al, Anal Biochem, 1994, 218, 112-117.
Kirchner et al, Techniques in Protein Chemistry VII (Marshak ed), 1996,
287-298.
Erdjument-Bromage et al, Protein Science, 1996, 3, 2435-2446.
Gharahdaghi et al, Anal Biochem, 1996, 223, 94-99.
Lui et al, Anal Biochem, 241, 1997, 156-166.
Hope this has been useful.
Joe
Joseph Fernandez
Associate Director
The Rockefeller university
Protein/DNA Technology Center
1230 York Ave. New York, NY 10021
Phone: (212)-327-8869
FAX : (212)-327-8620
email: fernaj@rockvax.rockefeller.edu
Lab Web Page: http:\\pdtc.rockefeller.edu