By Huaping Chen, Yuanyuan Li, Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)
The moment variation of Biological getting older: equipment and Protocols expands upon the former version with present, designated, important and promising tools at present to be had to review getting older. With new chapters on protocols that aspect getting older cellphone tradition in addition to many extra modern methods comparable to nuclear move, microarray and proteomics applied sciences. Written within the hugely winning Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and key pointers on troubleshooting and warding off recognized pitfalls.
Authoritative and functional, Biological getting older: tools and Protocols, moment Edition information the newest breakthroughs in know-how that experience served to improve the examine of aging.
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Additional info for Biological Aging: Methods and Protocols
Middle panel: Reanalysis of MRC-5 sorted in the PROL gate. Right panel: Reanalysis of MRC-5 sorted in the SES gate. (b) Majority of SES cells are positive for Sen-β-Gal. X staining, telomere-associated DNA damage, and BrdU incorporation [11, 28, 29] (see Fig. 1). 40 Graeme Hewitt et al. 1. Cells in the exponential growth phase are trypsinized and collected in DMEM plus 10 % FCS and immediately used for analysis and sorting in a FACS sorter (see Note 9). 2. Autofluorescence of unfixed cells is measured in FL1 (logarithmic).
Analysis of Biomarkers of Caloric Restriction in Aging Cells 27 4. 0 and develops blue staining only in senescent cells [19, 20]. SA-β-gal activity is routinely detected cytochemically, manually discriminating negative from positive cells by light microscopy. Flow cytometry-based SA-β-gal assay yields better differentiation to quantitatively calculate positivestained fibroblasts as compared to the routine cytochemicalbased method . 5. Senescent cells show striking changes in gene expression.
Analysis of Biomarkers of Caloric Restriction in Aging Cells 23 Fig. 1 Cellular senescence is determined by the senescence β-galactosidase (SA-β-gal) assay. Human WI-38 fibroblasts at a proliferating state (young, a) and senescence (old, b) are subjected to SA-β-gal staining and photographed. Blue staining cells are SA-β-gal positive or senescent cells. Magnification, Â100 4. Determine cellular replicative senescence at the end of late passage in the cellular lifespan by morphological changes (see Note 3) and senescence-associated β-galactosidase (SA-βGal) activity assay.
Biological Aging: Methods and Protocols by Huaping Chen, Yuanyuan Li, Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)