20S proteasome activity - 20S proteasome activity is determined via detection of fluorophores released during cleavage. Three proteasome activities are measured, the chymotrypsin-like activity, trypsin-activity (hydrophilic) and caspase-activity (acidic)

Beta-galactosidase assay - Official Campisi lab protocol for detecting senescence-associated endogenous beta-galactosidase activity. A flow cytometry protocol adapted from Kurz et al. is included

Catalase activity - Amplex® Red Catalase Assay kit from Molecular Probes. Instrument settings for the PolarStar Optima fluorometer from BMG Labtech is included

Cell culture guidelines - Cell splitting, passage numbers and aging cell terminology. Includes information on commonly used cell culture media, flasks, wells and incubator conditions

Cell cycle analysis - The DNA is stained with propidium iodide for DNA content determination by flow cytometry. The propidium iodide intercalates into the major groove of double-stranded DNA and produces a highly fluorescent adduct that can be excited at 488 nm with a broad emission centred around 600 nm

ELISA 96-well microplate - Conventional ELISA protocol (Enzyme-linked immunosorbent assay)

FACS Calibur - Includes information on instrumental aspects of flow cytometry

FACS Vantage cell sorting - A protocol for cell sorting by DNA content. The Vybrant® DyeCycle™ Orange Stain is used for DNA staining. The protocol also includes a short introduction to cell sorting technology

Flow cytometric detection of intracellular antigens - Cells are fixed in 0.25% paraformaldehyde and resuspended in fridge cold 70% methanol. The cell membrane is permeabilized with 0.25% Triton® X-100. The intracellular antigen is subsequently stained with a specific antibody solution containing a fluorochrome conjugated antibody

Immunofluorescence staining - The cells are grown in slide flasks in order to fix and stain the monolayers for conventional confocal microscope evaluation

MTT assay - Yellow 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) is reduced to purple formazan crystals in the mitochondria of living cells. The crystal development depends upon the integrity of the mitochondria and hence to some extent cell viability

Protein Expression & Purification - Protein expression in Hb2151 bacteria infected with phagemids and subsequent His-tag purification

SOD activity - Superoxide Dismutase Activity Assay kit from BioVision. The assay is measuring the amount of water-soluble formazan produced from the reduction by superoxide anion. The rate of the reduction of formazan with a superoxide anion is linearly related to the xanthine oxidase (XO) activity. The inhibition of formazan production in this assay is equal to the SOD activity

Splitting cells - Protocol for cell splitting in practice

Western Blotting - Whole WB procedure including gel electrophoresis and electroblotting