Dear Theresa,, , I would not advice to use thermofluor with membrane proteins as thefluorescent dye used in the assay binds to exposed hydrophobic patches of unfolding proteins. Therefore the presence of detergents would cause much unwanted fluorescence which would likely swamp your signal of interest. However, I have never personally tried it with a membrane protein .... . Best,, , D ________________________________________ From: Theresa Hsu [theresah...@live.com] Sent: 12 April 2014 09:38 To: ccp4bb Subject: [ccp4bb] Thermofluor assay
Dear all Does anyone has experience with Thermofluor assay to find the substrate transported/binding by a membrane protein? My protein does not have any similar structures and the substrate suggested by sequence analysis is not being transported in proteoliposome. I know ITC is good but I am looking for a more high-throughput way. Thank you. -- This e-mail and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorised recipient of the addressee please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to the e-mail. Any opinions expressed within this e-mail are those of the individual and not necessarily of Diamond Light Source Ltd. Diamond Light Source Ltd. cannot guarantee that this e-mail or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. Diamond Light Source Limited (company no. 4375679). Registered in England and Wales with its registered office at Diamond House, Harwell Science and Innovation Campus, Didcot, Oxfordshire, OX11 0DE, United Kingdom
