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Compatible detergents for proteomics


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4 replies to this topic

#1 Andreia Carvalho

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Posted 08 April 2011 - 03:53 AM

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

#2 mdfenko

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Posted 08 April 2011 - 11:10 AM

elute from what?
talent does what it can
genius does what it must
i do what i get paid to do

#3 Andreia Carvalho

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Posted 11 April 2011 - 12:58 AM

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
:)

#4 mdfenko

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Posted 11 April 2011 - 08:09 AM

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).
talent does what it can
genius does what it must
i do what i get paid to do

#5 johanski2

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Posted 12 April 2011 - 10:39 AM

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.

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






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