Jacob, You may also try DARTS (drug affinity responsive target stability). In this method, small molecule (or protein) binding leads to specific protection of its target protein from protease digestion. It is completely label free, and appears to be able to detect very low affinity interactions (high microM for small molecules). We'd be happy to send detailed protocols if you are interested.
http://www.pnas.org/content/106/51/21984.full Best, Jing -- Jing Huang, Ph.D. Associate Professor UCLA Department of Molecular & Medical Pharmacology 310-825-4329 jinghuang at mednet dot ucla dot edu http://labs.pharmacology.ucla.edu/huanglab/ ________________________________________ From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] On Behalf Of Jacob Keller [j-kell...@fsm.northwestern.edu] Sent: Monday, September 05, 2011 10:10 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Trying to "digest" PISA results NMR...take that! JPK 2011/9/5 Andreas Förster <docandr...@gmail.com>: > AUC ! > > > Andreas > > > > On 05/09/2011 6:00, Jacob Keller wrote: >> >> mea culpa! How about FRET? >> >> JPK >> >> On Mon, Sep 5, 2011 at 11:39 AM, Bosch, Juergen<jubo...@jhsph.edu> wrote: >>> >>> Hi Jacob, >>> you forgot cross-linking to stabilize a weak complex and verify that it >>> exists. >>> Jürgen >>> On Sep 5, 2011, at 11:45 AM, Jacob Keller wrote: >>> >>> Well, I guess I have always been curious what is the gold standard >>> here--perhaps SEC, ITC, SPR, pulldowns? What if SEC shows a >>> polydisperse sample with weak oligomerization, or SPR a very weak >>> binding constant? Do we then revert to a functional assay? Or what if >>> the functional assay does not show anything, but the binding constant >>> is really strong? Or vice versa, the binding is completely >>> undetectable, but the functional assay shows something? >>> >>> JPK >>> > > > -- > Andreas Förster, Research Associate > Paul Freemont & Xiaodong Zhang Labs > Department of Biochemistry, Imperial College London > http://www.msf.bio.ic.ac.uk > -- ******************************************* Jacob Pearson Keller Northwestern University Medical Scientist Training Program cel: 773.608.9185 email: j-kell...@northwestern.edu ******************************************* IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer.
