Brian,
In thinking about a proteomic project comparing tumors at different stages of development I came
across this reference:
Emmert-Buck, M.R. et at, Molecular Carcinogenesis 27:158-165 (2000)
where laser capture microdissection (LCM) was used to recover specific cell populations (50,000
cells per case) and recovered proteins were analyzed by 2D. Their method used AEBSF (the non toxic
serine protease inhibitor, 2 mM) in the staining baths. They lysed the cells by adding the LCM cap
directly into an eppendorf with IEF solution.
7000 LCM transfer pulses were needed to get their 50,000 cells and they approximated being able to
see only abundant (>50,000 copies per cell) proteins.
Linda
Big Apple Minute: Grey and dreary but the temperature is my favorite (60's F). My soon to be 16 year
old is so busy with studying for SAT II's and Reagents, writing a paper on Tienamin Square and
finishing the Red Cross Lifesaving Course that she doesn't want to think of planning her sweet
birthday.
Linda Siconolfi-Baez
Laboratory for Macromolecular Analysis
Albert Einstein College of Medicine
Ullman 405
1300 Morris Park Avenue
Bronx, N.Y.10461
718-430-2864
lsiconol@aecom.yu.edu
http://www.bioc.aecom.yu.edu/labs/angellab/
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