Protocol Online logo
Top : Forum Archives: : Protein and Proteomics

Solubilisation of proteins out of membranes - Which detergent at what concentration (Jan/24/2008 )

I am going to try to solubilise a recombinant protein out of membranes and I want to test different detergents for this porpose. Unfortunatelly I do not the the optimal condotions for each detergent (concentration, buffer, duration for sulobilisation, temperature, etc.)

Has anybody used some of the detergents for this purpuse and can help me out on this?

The detergents are

Triton X100
Tween20
Nonidet P40 (NP40)
Saponin (quillaya)
n-Octyl-beta-D-glucopyranoside (OGP)
CHAPS
SDS
Sodium-deoxycolate

I tried to firgure it out myself, but there does not seem to be a common role for the concentartion e.g. OGP is used at concentrations just above its CMC (30 mM) whereas NP40 is used in vast excess (0.5-2%).

-Dukes-

U absolutely right. no free cake.

u will have to test ur self which is the more suitable detergent and to optimize its concentration. u can also use

different protocol on the net.

what u should look for is Compatibility of reagents with ur purification system, meaning if its still works with high

concentration of those reagents.

Bye.

-amtash-