Hello Everybody,
I am working on a enzyme and its substrate. The enzyme is in the form of a membrane preparation (Sf9 ER fraction) and is resuspended in Hepes-NaOH (pH = 7.0) after ultracentrifugation. The substrate is expressed in E. coli, purified by Ni-NTA chromatography, and finally concentrated and resuspended in Hepes-NaOH.
I wanted to preserve both the enzyme and substrate at -80 degrees, I wanted to know if it is necessary to add glycerol as a cryoprotectant. The reason why I am asking this is that when I want to actually used it for the assay, the glyecrol might inhibit the assay?
Regards,
Kaushik
Hi Cheung,
Thank you very much for your reply.
What percentage or molarity of sucrose is generally used?
Will not affect my enzyme assay?
Regards,
Kaushik
Dear Stanley,
Thank you very much for your reply and the links.
I will go through them.
Best regards,
Kaushik
For proteins, on average, trehalose is more efficient than sucrose as cryoprotectant. As to how much to add - one must try that. It varies from protein to protein. The best long-term storage is frequently obtained by freeze-drying actually. Some proteins do not like it though.
- Badges
- Science topic
- Similar topics
- Cell Biology
- Methods