By Trygve O. Tollefsbol
This publication investigates a few of the strategies 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 equipment and protocols for those new options as well as its insurance of verified methods. Researchers looking new know-how and strategies will locate this quantity of super profit as they circulation in the direction of new instructions.
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Extra resources for Biological Aging: Methods and Protocols (Methods in Molecular Biology Vol 371)
Wash with PBS twice after blue color is developed. 5% of cold Triton X-100 in PBS at 4°C. 5% BSA in PBS for 20 min (see Note 4). 5% BSA either for 2 h at room temperature or overnight at 4°C (see Note 4). Wash three times with PBS, 10 min each. 5% BSA for 1 h at room temperature. Wash three times with PBS, 10 min each. Mount cells in mounting medium containing DAPI. Blue-colored staining for SA-β-gal activity is recognized well by bright-field microscopy and immunostaining is detected by epifluorescence.
5. Kodak D-19 Developer (Kodak). 6. Kodak Rapid-Fix (Kodak). 7. 0 (74 mM NaH2PO4, 9 mM Na2HPO4). 5. Immunostaining 1. 2. 3. 4. 5. 6. 7. Lab-Tek II Chamber Slide II (Nunc, Rochester, NY). 7% formaldehyde (see Note 1). 5% Triton in PBS. 5% BSA in PBS (see Note 4). 5% BSA in PBS (see Note 4). Secondary antibody (Bio-Rad, Hercules, CA). Mounting medium: VectaShield containing DAPI (4′,6′-diamidino-2-phenylindole; Vector Laboratories, Burlingame, CA). 3. 1. SA-β-Gal Staining for Adherent Cultured Cells 1.
1997) Telomere maintenance without telomerase. Chromosoma 106, 63–69. 8. Hayflick, L. and Moorhead, P. S. (1961) The serial cultivation of human diploid cell strains. Exp. Cell Res. 25, 585–621. 9. Cristofalo, V. , and Allen, R. G. (2003) Cell Senescence: An evaluation of replicative senescence in culture as a model for cell aging in situ, commentary and author’s response to commentary. J. Gerontol. 58A, 776–781. 10. Martin, G. , Sprague, C. , and Epstein, C. J. (1970) Replicative life-span of cultivated human cells.