Compatible detergents for proteomics - (Apr/08/2011 )

Hello everybody,

I am planning to perform proteomics, but I am having problems in the elution of my proteins. Does anayone know a detergent compatible with proteomics that I can use for the elution of the proteins that may increase the yield of the eluted proteins?

Thanks in advance,
Andreia

elute from what?

I want to elute from a agarose resine, but since I want to perform proteomics, i cannot use denaturating conditions. Do you know any detergent compatible with proteomics?

Thanks

protein A-agarose?

do you elute with low or high pH?

if low then you can try high pH, if high then try low pH.

i'm not sure that any detergent will help (maybe some tween-20).

If by "proteomics" you mean that you will be digesting the agarose eluate for analysis by LC-MS/MS, then you will want to use a MS-compatible detergent. The problem is that most detergents are not MS-compatible. Rapigest (sold by Waters) and sodium deoxycholate (SDC) have been used in our lab w/ much success, and you can find quite a bit about them in the literature regarding their use in proteomic experiments. Most other detergents are OK for solubilizing proteins, and many actually allow for more efficient digestion, but the problem comes when you try to remove them prior to LC-MS/MS (1, they make a mess of C18 chromatography, and 2. they often result in ion suppression in the MS).

With Rapigest or SDC, you can do a simple acid/solvent cleanup (see publications), and then carry on w/ your MS analysis.

I would not use detergents such as tween, or NP-40. SDS can be used, but it's a pain to clean up.

Andreia Carvalho on Fri Apr 8 11:53:03 2011 said: