Hi all,

Thanks in advance for any guidance here; I'm running a GTPase assay on recombinant human dynamin-related protein 1 (https://www.origene.com/catalog/proteins/recombinant-proteins/tp321708/drp1-dnm1l-nm_012062-human-recombinant-protein) and I had the protein off-ice (noob mistake) for ~15 minutes before adding it to my reaction samples. The protocol that my lab uses, which has produced results in the past, has a 30 min incubation period with GTP @ 37˚C. Given that the protein being in a liquid environment is baked into the protocol, and the test I ran today represented the end of the protein's first freeze-thaw cycle, is it likely that it survived 15 min at room temp? The assay came back negative & I'm trying to troubleshoot. Also, the buffer that the protein came in is 25 mM Tris-HCl, pH 7.3, 100 mM glycine, 10% glycerol.

Cheers!