Biological Aging: Methods and Protocols by Huaping Chen, Yuanyuan Li, Trygve O. Tollefsbol (auth.),

By Huaping Chen, Yuanyuan Li, Trygve O. Tollefsbol (auth.), Trygve O. Tollefsbol (eds.)

The moment version of Biological getting older: equipment and Protocols expands upon the former variation with present, precise, necessary and promising equipment at the moment to be had to review getting older. With new chapters on protocols that element getting older telephone tradition in addition to many extra modern techniques reminiscent of nuclear move, microarray and proteomics applied sciences. Written within the hugely winning Methods in Molecular Biology sequence structure, chapters contain introductions to their respective themes, lists of the required fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and key pointers on troubleshooting and heading off identified pitfalls.

Authoritative and functional, Biological getting older: tools and Protocols, moment Edition info the latest breakthroughs in expertise that experience served to increase the learn of aging.

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Apoptotic cells and debris are excluded from analysis. Mean fluorescence is determined in FL3 (logarithmic) using a FSC/FL3 scatter plot. Median autofluorescence of an unstained control is also analyzed. Similarly to MitoSOX, sorting gates are defined in FL3 (logarithmic), conveniently in a FSC/FL3 dotplot. Use the left and right quartiles with respect to FL3 to sort cells with highest (high FL3) and lowest (low FL3) superoxide levels. 4 Notes 1. Magnetic affinity cell sorting (MACS) can also be used to sort cells.

Compare alterations of these parameters in control and CR cells at different PDs during aging progression. 4 Age-Related DNA Damage Analysis 1. 2, step 3 followed by the routine protocols. Analysis of Biomarkers of Caloric Restriction in Aging Cells 25 Table 2 Selective CR-related genes Expression change Gene Function in CR References SIRT1 NAD-dependent histone deacetylase; activation of SIRT1 is seen in different animal organs affected by CR Increased expression [14, 29] Foxo Forkhead transcription factors; controls various biological functions; involves CR-related longevity Decreased expression [29, 30] PGC-1α Regulates mitochondrial function and glucose homeostasis during CR Increased expression [29, 31] PPAR-γ Insulin sensitizer; increase insulin sensitivity during CR Increased expression [28, 32] Glucose-6phosphate Involve energy metabolism such as glycolysis Increased expression [28] Hsp105 Heat shock protein; stress-related gene Decreased expression [28] 2.

For example, upregulation of SIRT1, an NAD-dependent histone deacetylase, has been confirmed to correlate with CRinduced longevity [14, 29]. Other known CR-related genes can be detected to determine the efficacy of this in vitro CR system [28–32]. 28 Yuanyuan Li and Trygve O. Tollefsbol Acknowledgments This work was supported by grants from the American Institute for Cancer Research (AICR), the National Cancer Institute (CA 129415), Susan G. Komen for the Cure, and the American Cancer Society Award (IRG-60-001-47).

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