In screening detergents that solubilize my protein of interest from IM preps in E. coli (after overexpression from a pET vector), I found that Na-deoxycholate and lauryl sarcosinate are the most effective.
However, these detergents are not compatible with IMAC so they're not ideal for downstream purification after solubilization (POI is 6XHis-tagged)
Are there other similar detergents to these I can screen that might be more compatible with IMAC? I have already tried UMNG, C-Hega-8, MEGA-8, CyMal 1-4, CyGlu-4 and none of these solubilize my protein at 2%
Any other suggestions would also be welcome!
Thanks in advance
The detergents DM, LDAO, DDM Cy6 and FOS are suitable for solubilization of Presenilin. The Ni-NTA Membrane Protein Kit was used to screen for the most suitable detergent for solubilization of the human membrane protein Presenilin. You may try these too!
CHAPS and CHAPSO are zwitterionic detergents with a similar structure to the anionic detergents cholate and deoxycholate. Bio-Rad lists them as compatible with their Profinity IMAC resin at concentrations up to 5%.
Kara Hood SLS is pretty 'aggresive' as a detergent, we often use it as a positive control since it extracts pretty much everything, but it doesn't really keep the protein functional.
Regarding deoxycholate, you can try using CHAPS and CHAPS-O (rather at high concentrations), otherwise try using a mixture of detergents containing cholate since it does the work for you.
And you mentioned that these detergents are the most effective, but bear in mind that solubilizating everything is not always good : you may inactivate your protein etc., so even lower solubilization yields (60-80%) can be very good.
Dear Kara
Is it your protein a membrane protein?
Detergents ad DM, DDM, OG (which are non ionic detergents) are expensive but are often used for solubilization on membrane proteins.
In my blog ProteoCool ProteoCool i just upload a presentation
ProteoCool n°34: Inappropriate use of ultrafiltration for concentration and/or buffer exchange of membrane protein
and in the first part of it you can find some general consideration about detergents and their properties.
ciao
Manuele
Detergent Screening is always a annoying and difficult process when solubilizing your protein.
Just to giye you an idea how many are out there:
https://cube-biotech.com/products/ultrapure-detergents/?p=1
But as Manuele Martiniella pointed out. OG and DDM might be worth a try here:
https://cube-biotech.com/products/ultrapure-detergents/octylglucoside-og/
https://cube-biotech.com/products/ultrapure-detergents/dodecylmaltoside-ddm/
However may I suggest you to use an alternative to detergent solubilization, namely nanodiscs, maybe they are an option for you:
https://cube-biotech.com/us/what-are-nanodiscs
Can we use sodium lauryl sarcosine as an alternative for sodium deoxycholate for RIPA lysis buffer preparation.
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