Could anyone provine hints on how to extract and quantify protein from shoot samples of chestnut tree (Castanea sativa)? We have been trying to extract proteins from frozen leaf samples (between 200 and 500 mg) using 100 mM PK buffer with 1 mM EDTA, 8% glycerol, 1 mM PMSF, 5 mM ascorbate and PVPP, but unfortunately no protein was detectable using Bradford's method, even without diluting the supernatant. We have tried the exact same procedure with tomato and barley samples and protein levels were detectable and in normal ranges.
Thank you Rani P Ramachandran
!Although it is interesting to know that the authors had also had difficulty in extracting protein from chestnut tissues, I'm afraid the extraction methods tested in this paper are too strong and would affect the structure and function of proteins, disrupting enzyme activity, which we aim to evaluate..
- Badges
- Science topic
- Similar topics
- Filing