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Position: Discussion forums > Bioscience biotechniques discussion forums > Cell biology and histology methods forum
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caspase assay protocol for yeast
Posted by: beachbum (IP Hidden, New member, 1)
Date: March 7, 2006 03:40PM
Hi, I'm a high school student conducting a science fair project on the presence of the YCA1 caspase in P. pastoris. I have asked around but nobody seems to know how to determine the activity and presence of this caspase in yeast and upon searching for methods, I have come across flow cytometry, spectrophotometry, and fluorimetry techniques however, all seem to be suited for mammalian cells and there aren't any step by step procedures. Does anyone know of a good caspase assay protocol I can use to detect YCA1 activity in P. pastoris?
Additionally, I'm also trying to induce nitric oxide production by inducing the nitric oxide synthase enzyme in this yeast however, this again is usually done in mammalian cells and NO induction is achieved by addition of a wide variety of cytokines and lipopolysaccharide. Upon searching for literature, no results have come up regarding yeast sensitivity towards LPS addition and it is unlikely for LPS to cross the yeast cell wall. Do you know of a NO promoting drug that I could use in yeast? Thankyou so much for your time! Amy.
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