Hi everyone, sorry for this none CCP4 question,
but I am working on some tough proteins which easily got aggregated
after purification, I am thinking of using some detergents rightnow,
however, I am a newbie in this field, could you please give me some advice on
the usage of detergents?
here is some basic information about the protein:
1. This protein is as big as 90KD.
2. The productivity of the protein is very low, and the
homogeneity of the protein is not good. Most of the proteins come out of the
column at void volumn, and the rest of the proteins forms 3-4 peaks, and each of
them features low peak height.
3. I have already tried different truncates of the protein and
homologous proteins from other species, up to now the protein on
my hand is the best of them.
4. I have stocks of the following detergents:n-Octyl-b-D, n-Decyl-b-D,
FOS-Choline-12, n-Dodecyl-b-D, Cymal-5, CHAPS, Tween20,
Triton-X100.
my question is:
1. would you please recommend some other detergents to
try?
2. should I try different concentration of the detergent and how? right now
I just tried the concentration equals the CMC, some of the
detergents do improve the homogeneity of the
protein, however, once I concentrate the protein for crystallization, the
detergent get concentrated too, and as you know that is very bad for
crystallization.
3. would you please recommend me some literature to resort to (about
the same situation as mine or about the use of detergents)?
4. any other suggestions or comments about how to improve the quality of
the protein.
Your suggestions and comments will help me a lot and will be highly
appreciated.
Many thanks to all of you!
Bei
2010-09-07
joybeiyang