Dear All, I am in a situation, almost for the first time within my limited experience, that deglycosylation might be necessary to obtain crystal. So, I thought of tapping to vast experience of CCP4BBers, while I am searching literature.
I have protein that has been expressed in HEK293 cells, secreted into media, purified over IMAC and SEC columns. Crystallization-screens with its binding partners (they form good complexes based on analytical SEC) have not produced any useful hits (whereas complexes with related proteins worked well). So, I plan to re-try complex formation and \crystallization screen after deglysosylation. My question is: In practice, Does a kit (for example here: https://www.sigmaaldrich.com/catalog/product/SIGMA/NDEGLY?lang=en®ion=US) containing Endo F1, F2, F3 be sufficient or should this be tried in combination with PNGase (which requires desaturating conditions)?!! Many Thanks in advance for your suggestions, and reference. Best Wishes, Partha ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
