Dear Ammrl group,
I received a number of good social and technical suggestions for acquiring 15N-HSQC on Broadband probe. After reading all your suggestions, I run 15N-CIGAR (attach) instead of 15N-HSQC to test my cable connection. The sample I used was 2% 15N-benzamide. I also auto calibrated the probe for 15N nucleus. The tn='H1' and dn='N15', changed the ¼ loop and put in 15N filter. I am able to see signals that means the connection is fine. I should be able to get 15N-HSQC for small organic molecules but the protein sample is difficult. I couldn't get any signal.
Many of you suggested using HCN and Bio-pack combination for 15N enriched proteins in water. I agree, I am able to get 2D 1H-15N correction using gNhsqc within 5 min. My challenge is convincing this particular faculty to use HCN and biopack. I may need to show him all the S/N ratio, spectra, and acquisition time to convince this senior faculty, sticking to the old method. All the information are helpful and thank you.
Hla Win-Piazza, Ph.D.
Organic Chemistry Lab Coordinator
Western Washington University
Department of Chemistry
Received on Thu Mar 06 2014 - 11:10:41 MST