Flow cytometiy

Figure 2. Detection of 02- in BAEC upon treatment with CsA. Left representative experiment of superoxide detection in supernatants of BAEC measured by electron spin resonance (ESR) using the superoxide-sensitive spin trap DMPO. The superoxide generator DMNQ was used as a positive control. Right flow cytometiy detection of 02 with die probe DHE. BAEC were preincubated for 1 hour with 5 pM DHE and then incubated for 2 hours with the indicated doses of CsA. Data are represented as mean intracellular fluorescence of ethidium (oxidized fluorescent form of DHE) (n = 7). p < 0.05 vs CsA vehicle (0 pM CsA).

Methods for cell and sub-cellular fractionation include immuno-isolation, electromigration e.g. free flow electrophoresis), flow cytometiy, density gradient isolation of organelles and sequential extraction. The use of some of these approaches to investigate the mitochondrial and myofibrillar sub-proteomes of the heart will be described in Sections 3.4 and 3.6 respectively. A particular problem in proteomic analysis of the heart is the diversity of cell types that are present. Proteomic profiles of total myocardial lysates are dominated by the proteins present in cardiac myocytes, but such samples... 

The choice of label will depend on the application envisaged. Enzymes are widely applicable they are used in assays, such as ELISAs, and for detection of antigen blotted or dotted on membranes or embedded in tissue sections. Enzyme labels have also be used for the location of antigen in electron microscope sections but gold labelled antibodies (Chapter 11) are now extensively employed for this purpose. Antibodies labelled with a fluorescent molecule are used in assays and for the detection of antigens in tissue sections and also for flow cytometiy and fluorescence-activated cell sorting. 

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