By Huaping Chen, Yuanyuan Li, Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)
The moment version of Biological getting older: tools and Protocols expands upon the former version with present, unique, important and promising tools at present on hand to review getting older. With new chapters on protocols that aspect getting older mobilephone tradition in addition to many extra modern methods corresponding to nuclear move, microarray and proteomics applied sciences. Written within the hugely profitable Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective issues, lists of the required fabrics and reagents, step by step, simply reproducible laboratory protocols, and key tips about troubleshooting and averting recognized pitfalls.
Authoritative and sensible, Biological getting older: equipment and Protocols, moment Edition info the newest breakthroughs in know-how that experience served to improve the learn of aging.
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Extra resources for Biological Aging: Methods and Protocols
Tollefsbol Normal human fibroblasts Normal glucose level medium Age progression in normal cells Glucose restriction medium (CR) PDs calculation Age delay and lifespan extension in CR cells SA-b assay Analyze age-related oxidative stress and DNA damage Compare age- and CR-related differential gene expression Fig. 2 Schematic representation of the procedure for aging biomarker analyses in the CR in vitro model 4 Notes 1. Normal human diploid fibroblasts such as fetal lung fibroblasts, skin fibroblasts, and vascular fibroblasts with a finite replicative lifespan in vitro are the most commonly used cell types to manifest aging [16, 17].
89 ml DMSO to the 10 mg vial. As DHR123 is readily oxidized by air, the stock solution is immediately aliquoted under N2 atmosphere in 100 μl each and frozen at À20 C. 2. 5,50 ,6,60 -Tetrachloro-1,10 ,3,30 -tetraethylbenzimidazolylcarbocyanine iodide (JC-1) (Molecular Probes 5 mg, Cat# T-3168). 5 mg/mL in dimethylsulfoxide (DMSO) by adding 2 mL of DMSO to the 5 mg vial. To avoid repeated freeze/thaw cycles, aliquot the stock immediately into portions, each sufficient for one day of experimental work, and store them at À20 C until required for use.
Non-senescent cells cannot be obtained, and sorting will result in an enrichment of senescent cells, but not in the production of a “pure” population. 1 Materials Cell Culture 1. Dulbecco’s modified Eagle’s medium (DMEM) with 10 % fetal bovine serum, 1 % penicillin/streptomycin and 1 % glutamate. 2. 1Â phosphate-buffered saline [1Â PBS]. 3. Solution of trypsin–EDTA (5 g of trypsin, 2 g of EDTA per liter of distilled water). X in Fixed Cells 1. Paraformaldehyde (2 %). 2. 5 % Triton X-100 in phosphate-buffered saline [1Â PBS]).
Biological Aging: Methods and Protocols by Huaping Chen, Yuanyuan Li, Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)