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Dear NMR wikiers I would like to take c13 filter noesy experment ( this experment explore residues of the interface of two monomers of homodimer protein ).homodimer protein size is total 20 kd . could you please suggest which parameters should i consider more specificly while taking this experment to get best signal because i dont want to try on trial and error method. any knid of help is greatly appreciated. Thanking you in advance Regards sri |
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Same parameters as for a 20 kDa monomer? NOESY mixing time 50-100 ms, depending on how much spin-diffusion you want to risk. Or, is this the 90% deuterated sample that you have asked about before? If so, you could afford a longer mixing time, though clearly the sensitivity is going to compromised by the low level of protonation, though this will be less serious if you are looking at C-H <-> N-H NOES in H2O solution. [P.s. Basically your questions lack sufficient information to provide decent answers. For example, you say filter NOESY. Is this a mixture of protonated and deuterated or 12C and 13C chains?] No , it is not deuterated protein sample , protein monomer size is 10 kd and prepared sample by mixing C13 and C12 samples ,I mixed LB medium protein sample with C13 labelled sample to get intramoleculer NOEs .I am trying now NOESY mixing time is 150ms and geting lot NOE leakage. - sri (Jul 05 '11 at 00:37) my protein exist as always dimer. - sri (Jul 05 '11 at 10:56) |
