alternatives to triton-x and chaps? - (Sep/18/2008 )

hello,
can i have suggestions to alternatives rather than using triton-x or chaps. i would like to avoid using these as they are detergents and i want to analyze my proteins by mass spectrometry. what are good alternatives to these detergents?

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it depends of course of what you want to do, but freeze-thaw cycles will also lyse cells, as will sonication or using a French press ...

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You can usually run a desalting column before ESI-MS. For MALDI, this desalting "column" is a Zip Tip which can be used to remove salts and other small molecules (i.e. small detergents) from the protein sample. This would allow for you to use the detergent for whatever you want it for then remove it prior to MS analysis.

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I know you want to avoid detergents but still some detergents are very mild and are no harm. CHAPS is a mild detergent......not as strong as NP40 and Triton X100, Tween 20.....you can check the list of biological detergents on SIGMA. ...why is your protein in CHAPS or Triton anyway? are they cell lysates? in that case you can do what dpo suggested in his post.

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You can still use the detergents but you should run your samples in a sds-page gel before the MS....

Otherwise I did for the same pupose (MS/MS) for my samples in Low salt buffer (LSB)
I needed whole protein extraction method...so collected the cells in LSB, left them for 5 min at room temperature, then store them at -80 degrees for at least 1 hour. the cells will swell and burst....take the samples out of the freezer, thaw them (you can even sonicate them)...centriguge 10 min at top speed..take supernatant.

Or also you can use 6M urea + 2 M thiourea (be carefull with the protein quantification method you use...is high concentration of urea)

regards

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