RIPA buffer (Radio Immunoprecipitation Assay buffer)
150 mM sodium chloride
1.0% NP-40 or Triton X-100
0.5% sodium deoxycholate
0.1% SDS (sodium dodecyl sulphate)
50 mM Tris, pH 8.0
Ripa buffer is used for extraction of fungal protein but I have no sodium deoxycholate and Triton X-100. what can I use instead of them?
I think it is posible to replace Triton X-100 by Tween 80 and leave out sodium deoxycholat. There is a manual from Calbiochem: http://wolfson.huji.ac.il/purification/PDF/detergents/CALBIOCHEM-DetergentsIV.pdf
where you can check which Detergent is suitable
Thanks for the answer. I use Protein Purification Protocols Second Edition Edited by Paul Cutler(book) for the preparation of buffers.
I would not expect these to be equivalent- tween is a non-ionic detergent with a polysorbate moiety while sodium deoxycholate is an ionic detergent with a cholesterol moiety. I doubt the tween would be able to do much to solubilize membrane proteins (especially if it is a low MW version like tween 20). If you are searching for a substitute you might consider detergents from the CHAPS series which share a steroid moiety.
Triton X-100 and NP-40 are non-ionic like tween but tend to be more aggressive. They are inexpensive and widely available so you should try to seek them out. If you are unable to obtain them you might try one of the lower MW Brij varieties.
If you do proceed with substituting take note of the CMC (critical micellar concentration) of the substitute and adjust concentrations accordingly.
I usually do viral nucleocapsid purifications for whole genome sequencing and use both Sodium deoxycholate and NP 40 for cell lysis. As both Tween and NP-40 are non-ionic detergents you can use any of them (although I never used Tween for cell lysis), better is not to leave ionic detergent Sodium Deoxycholate(10 gm is max 30 USD).
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