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My Protocols

  • 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 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

  • Cleaning the Autoflex II ion source - Autoflex II ion source cleaning procedure

  • ClinProTools basic workflow - The software performs statistic analysis of peaks from different spectra. It can also run sophisticated mathematical and bioinformatic algorithms for pattern recognition model generation

  • ELISA 96-well microplate - Conventional ELISA protocol (Enzyme-Linked ImmunoSorbent Assay)

  • ESI-Q/TOF - Applied Biosystems Qstar XL and Elite mass spectrometers. Guidelines and information on extern calibration, syringe connection, collision energy adjustment, needle washing, LC-ESI interface handling, data acquisition, postprocessing and database search

  • 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

  • Glycogen content in liver homogenates - Glucose is measured before and after glycogen hydrolysis in a strong acid. The glycogen concentration is calculated by subtracting the free glucose from the glucose released by hydrolysis

  • Hepatic clinical-chemical analysis - Tissue homogenization, lipid extraction, lipid saponification, and ADVIA 1650 clinical chemical system from Siemens. Analysis of phospholipids, triglycerides, total fatty acids, glucose and glycogen

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

  • iTRAQ labeling of proteins - Protein content determination (BCA method), acetone protein extraction, disulfide bridge reduction, trypsination, iTRAQ labeling, column sample loading, freezdrying and sample storage

  • LC-MS/MS systems - Agilent 1100 Module Series with ChemStation software. Strong Cation Exchange (SCX), nano-RP-HPLC and capillary RP-HPLC

  • MALDI-TOF/TOF - Bruker Daltonics Autoflex II mass spectrometer. Guidelines and information on matrix preparation, acquisition modes (linear vs. reflecton mode), LIFT technology, FlexControl automation, FlexAnalysis, BioTools, WARP-LC software, instrument settings and quality control procedures

  • 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

  • ProteinBrowser results - Compound list information and protein/peptide table parameters. Explanation and interpretation of the output

  • ProteinPilot results - Applied Biosystems/MDS Sciex ProteinPilot. Explanation and interpretation of the output

  • 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

  • Thin layer preparation with HCCA matrix - Thin layer preparation on the AnchorChip with 800/384 geometry

  • Tissue, blood and milk sampling - Tissue samples, liver biopsies, blood and milk samples

  • Western Blotting - Whole WB procedure including gel electrophoresis and electroblotting