I have diphtheria toxin (sigma D0564), which is in the unnicked form.
The product info page says it has to be nicked by a protease and then treated with DTT to be active, but I assume this is only required for in vitro biochemical assays.
I would like to inject DT to deplete DTR expressing cells. From what I've read, people simply reconstitute the powder in water, aliquot and freeze in -80C (freeze thaw seems to significantly affect DT activity).
Essentially what I want to know is can I simply reconstitute in water or do I need to nick the DT as well with a protease such as trypsin, and then aliquot and freeze.
Much thanks,
Yong
Dear Sir. Concerning your issue about the preparation of the Diphtheria Toxin solution for in vivo depletion of specific DTR expressing cells . I think the following below links may help you in your analysis:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4194082/pdf/nihms572036.pdf
https://open.bu.edu/bitstream/handle/2144/16050/Lee_bu_0017N_11063.pdf?sequence=1
Thanks
Hi YongLin, I have the same question. Have you figure it out? And many people say that DT is very sensitive to freeze and thaw. Even one freeze-thaw cycle makes its activity gone. is it real? How you make your work solution? Thanks!
Lingling Xu
I am also using DT to abate hair cells in vivo. Have you figured out how to prepare the Diphtheria Toxin solution? Is nicking required? Thanks!