****************************************************************
R. Michael Garavito, Ph.D.
Professor of Biochemistry & Molecular Biology
513 Biochemistry Bldg.
Michigan State University
East Lansing, MI 48824-1319
Office: (517) 355-9724 Lab: (517) 353-9125
FAX: (517) 353-9334 Email: [log in to unmask]
****************************************************************
Hi Jose,
how do you know that those crystals were detergent and not protein? My impression is that it is really hard to crystallize DDM, and even harder for DM (solubilities > 20% in water). The easiest (?) way to check this may be to take some crystals, wash them well and run them out on a PAGE gel. If you don't see anything and you've taken enough crystals, then you're probably dealing with pure detergent crystals. As for your second point, you're right. For most low-cmc detergents the total detergent concentration will be substantially higher than reported, since a substantial amount is always bound to your protein. For 1 mM DDM, you would have only ~ 20 uM micelles, assuming an aggregation # of 50 (its higher). I don't think people measure the total detergent concentration in the end; for maltosides one could in principle do a Fehling's based assay to get the concentration.
Cheers, Bert
Bert van den Berg
University of Massachusetts Medical School
Program in Molecular Medicine
Biotech II, 373 Plantation Street, Suite 115
Worcester MA 01605
Phone: 508 856 1201 (office); 508 856 1211 (lab)
e-mail: [log in to unmask]
http://www.umassmed.edu/pmm/faculty/vandenberg.cfm
"Parveen Goyal" wrote:
> Hi All,
>
> I got some hexagonal crystals in one of my crystal condition. The protein is
> a membrane protein and contains 0.05% DDM. Has anybody seen DDM crysals
> and > if yes, how do they look like?
>
> thanks in advance
>
> Parveen Goyal
>