Download Apoptosis: A Practical Approach (Practical Approach Series) by George P. Studzinski PDF

By George P. Studzinski

This crucial textual content offers conceptual outlines and exact techniques for simple and complex experiences of mobilephone loss of life by way of apoptis. Chapters at the attractiveness of apoptis as individual from neurosis and nonspecific mobilephone DNA harm are through a scientific exam of the verified and the relevant novel methodologies used by prime laboratories carrying out examine on apoptis. a wide selection of techniques are supplied, allowing readers to take part in state-of-the-art research.

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These include the development of antibodies to single-strand DNA breaks (30); biotinlabelled, hairpin oligonucleotides that bind to double-stranded DNA breaks (31) and antibodies to caspase-cleavage products, such as the carboxyl terminus fragment of actin (28). One of the most common techniques used is the detection of phosphatidylserine in the outer leaflet of the plasma membrane of apoptotic cells, using labelled annexin V. This technique is the subject of Chapter 7. Here, it is sufficient to say that the technique can be used in conjunction with both light and electron microscopy.

3. Give the slides 10 dips in 95% ethanol. Repeat. 4. Transfer to absolute ethanol for 10 min. 5. Treat as for Protocol 8, steps 7 and 8. Thionin may be reused many times before staining intensity is impaired. Methanol (80%) may also be reused, until it becomes too discoloured. 2 Stains for fluorescence microscopy Fluorescent nuclear counterstains are most appropriate for use with cultured cell systems, and when fluorescent detection methods are being used in parallel for immunohistochemistry. The most commonly used are 4',6-diamidino2-phenylindole (DAPI), Hoechst 33258 and 33342, acridine orange, and propidium iodide.

Duodenum, jejunum, upper ileum, lower ileum, caecum, mid-colon/rectum. 3. Cut into 1 cm lengths. 4. 'Bundle' together up to 10 pieces of intestine: (a) cut an 8-10 cm length of micropore tape and form a loop by sticking the two ends together, (b) lay the pieces of intestine parallel to each other within the loop and bind together with micropore tape, (c) cut a 4-5 mm thick cross-sectional disc from the taped bundle. 30 2: Morphological recognition of apoptosis 5. Process these smaller bundles for histology, as described in Protocol 3.

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