Apoptosis: Techniques and Protocols (Neuromethods) by Andréa C. LeBlanc PDF

By Andréa C. LeBlanc

ISBN-10: 1588290123

ISBN-13: 9781588290120

ISBN-10: 1592591884

ISBN-13: 9781592591886

During this revised and increased moment version, pro specialists describe in step by step aspect their most sensible state of the art thoughts for learning neuronal cellphone demise. those without difficulty reproducible tools remedy a wide selection of analysis difficulties, together with the detection of the main proteins fascinated by neuronal apoptosis (Bax protein, cytochrome c, and caspases), the direct evaluate of the function of pro-apoptotic proteins in neurons through viral infections and microinjections, and the detection of pro-apoptotic proteins in situ. There also are hands-on equipment for the learn of apoptosis mechanisms in neuronal booths, for learning synaptosis, and for constructing gene expression profiles in neurodegenerative mind tissues by utilizing DNA microarrays.

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4. , and Vassalli, P. (1997) An early and massive wave of germinal cell apoptosis is required for the development of functional spermatogenesis. EMBO J. 16, 2262–2270. 5. , Mai, J. , Mossakowski, M. , and Reed, J. C. (1995) Upregulation of Bax protein levels in neurons following cerebral ischemia. J. Neurosci. 15, 6364–6376. 6. , Nitahara, J. , et al. (1996) Programmed myocyte cell death affects the viable myocardium after infarction in rats. Exp. Cell Res. 226, 316–327. 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 Bax Localization and Purification 19 7.

If you get air, then try to remove them by gently pushing on the cover slip with blunt tweezers. It is often best to use a little extra mounting medium and remove the excess at the edge of the cover glass with a 10-µL pipet. 5. Seal the cover glass in place with CytosealXYL. 6. 17-mm cover glass onto the slide. 7. Store fluorescently labeled samples in the dark at 4ºC. 11. Mitochondrial Co-localization Co-localization of cytochrome-c, Bcl-2, Bax, or other Bcl-2 family members with mitochondria can be done in fixed cells using Mito- 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 Cytochrome-c Release 27 tracker Green (Molecular Probes, Eugene, OR), which does not depend on inner mitochondrial membrane (IMM) potential (∆Ψm) for mitochondrial localization.

2. Fluorescence will fade. 3. Reagents and Solutions See Table 1. 4. Antibodies for Immunostaining See Table 2. 5. Paraformaldehyde Fixative 1. 0 g paraformaldehyde in ~60 mL H 2O with one drop of 10 N NaOH. 2. Warm to no more than 60ºC, on a heating stir plate. 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 24 Ethell and Green Table 1 Reagents and Solutions for Immunostaining Reagent Catalog number Vendor Bovine serum albumin (BSA) Chambered cover glass Cover slips CytosealXYL BP1605-100 155409 12-545-82 8312-4 DAPI Dimethyl sulfoxide (DMSO) Normal goat serum (NGS) Paraformaldehyde 10X phosphate-buffered saline (PBS) Tween-20 Vectashield mounting medium D8417 D2650 G9023 T353-500 70011-044 Vectashield + DAPI H-1200 Fisher Nalge-NUNC Fisher Stephens Scientific (Kalamazoo, MI) Sigma Sigma Sigma (St.

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Apoptosis: Techniques and Protocols (Neuromethods) by Andréa C. LeBlanc


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