Read e-book online Biological Aging: Methods and Protocols PDF
By Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)
Biological getting older: tools and Protocols investigates some of the techniques which are suffering from the age of an organism. a number of new instruments for the research of organic getting older were brought lately, and this quantity offers tools and protocols for those new recommendations as well as its insurance of confirmed approaches. The editors have rigorously chosen merely these subject matters which are thought of mainstays of the sector or are exhibiting promise in revolutionizing this really new technology. the 3 major parts of concentration during this state of the art compendium of organic getting older learn are: tools which are uncomplicated to figuring out the basic mechanisms of mobile getting older; ideas used to intrude within the getting older technique; and methods to reading the various molecular procedures of organic getting older. Researchers looking new expertise and strategies will locate this quantity of great profit as they circulation in the direction of new instructions within the interesting and increasing box of organic getting older.
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Extra info for Biological Aging: Methods and Protocols
4. The pellet is resuspended in 3 mL of serum-free DMEM and is immediately used for analysis and sorting (Note 12). 5. ). Sorting gates are defined in Fl1 (logarithmic), conveniently in a FSC/FL1 dotplot. Use the left and right quartiles with respect to FL1 to sort cells with highest (high FL1) and lowest (low FL1) peroxide levels. 2. JC-1 STAINING FOR DETERMINING MMP (NOTE 13) 1. 1 × 105 cells are resuspended in 500 μL of RPMI 1640 phenol-free without supplements (Sigma) and 1 μL of JC-1 stock solution is added to give a final concentration of 1 μg/mL.
2. With the exception of BJ fibroblasts and other primary cell cultures, 1–2 min submerged in trypsin at room temperature or 37°C should be sufficient to disperse cells. BJs and other difficult cell types may require 5–15 min of incubation at 37°C, with occasional checks under an inverted microscope to ensure that cells are detached. To avoid clumping of cells with fibroblast-like morphology, do not agitate the cells by hitting or shaking the flask/culture vessel. Continue to place cells at 37°C until they are no longer adherent or until 15 mins have passed (extended trypsin exposure damages cells).
Wang, R. J. (1976) Effect of room fluorescent light on the deterioration of tissue culture medium. In Vitro 12, 19–22. 14. Bradley, M. O. and Sharkley, N. A. (1976) Mutagenicity and toxicity of visible fluorescent light to cultured mammalian cells. Nature 266, 724–727. 15. Bradley, M. , Hsu, I. , and Harris, C. C. (1979) Relationship between sister chromatid exchange and mutagenicity, toxicity and DNA damage. Nature 282, 318–320. 16. , Chen, Q. , and Toussaint, O. (2000) Cell cycle regulation in H2O2 induced premature senescence of human diploid fibroblasts and regulatory control exerted by the papilloma virus E6 and E7 proteins.
Biological Aging: Methods and Protocols by Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)