I can second this. Whether or not you get a signal with DSF depends on your protein, not on the type of ligand. We also use Sypro orange. Herman
________________________________ From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Bosch, Juergen Sent: Thursday, June 28, 2012 1:42 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] off topic: protein peptide interaction using DSF Yes add peptide in different concentrations to your protein and run the assay* Yes, or Sypro Orange *Rule of thumb 20 kDa protein use 1 mg/ml 50 kDa protein use 0.5 mg/ml also have a peptide which should not interact with your protein as control Jürgen On Jun 28, 2012, at 7:34 AM, rashmi panigrahi wrote: Hi all, Has anyone performed protein peptide interaction experiments using flurophore on a real time PCR machine? Please advice me on how the experiment is performed. Can I use ANS? with regards -- rashmi ...................... Jürgen Bosch Johns Hopkins University Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Office: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-2926 http://web.mac.com/bosch_lab/