Download Adipose Tissue Protocols by Gérard Ailhaud PDF

By Gérard Ailhaud

ISBN-10: 0896037479

ISBN-13: 9780896037472

ISBN-10: 1592592317

ISBN-13: 9781592592319

Adipose tissue is now famous as a widely-dispersed secretory organ that performs an important function in weight problems. Ailhaud (U. de Nice-Sophia Antipolis, great, France) and a staff of eu and US laboratory specialists and clinicians aspect the type, key ideas hired, and their power difficulties, within the in vivo and in vitro examine of brown and white fats tissues and cells from the dietary, physiological, and molecular degrees. significant therapy is accorded to the isolation, subcellular fractionation, and gene move into mature adipocytes, in addition to metabolic points. Illustrations contain microscopy research effects, and a guy donning a conveyable detector for measuring adipose tissue blood stream.

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Extra resources for Adipose Tissue Protocols

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8. 1% OsO4 in PBS, for 50 h at 20°C or for 3 h at 45°C. 9. Multiple rinses in PBS. Morphologic Techniques for BAT and WAT 41 10. Dehydration in ascending EtOH concentrations and critical-point drying. 11. 5–2 nm) with a sputter coater. This coating is thinner than that used for conventional SEM observation, and allows study of cells in their finest ultrastructural detail, but requires such careful tissue handling that, for instance, specimens cannot even be moved from one laboratory to another.

For the latter, the material to be sectioned must be able to withstand the heat generated by the electron beam; therefore, specimens must be embedded in a hard epoxy resin. After fixation, specimens are dehydrated in increasing concentrations of dehydrating agents, such as alcohol. This is necessary because the embedding medium is not water soluble. After dehydration, specimens are infiltrated with a liquid embedding medium (a plastic monomer), which is then polymerized to produce a solid block using the epon-araldite mixture (20).

For ~1 mL packed cells, use 40 mL of fixative solution, with gentle shaking, on a rocker platform (see Note 2). Use 15- or 50-mL polypropylene tubes (see Note 3). 2. 4, containing 50 mM glycine, to quench aldehyde-induced nonspecific binding sites. 3. Permeabilize and block nonspecific binding sites with B1 for 45 min at RT. 5-mL polypropylene screw-cap tubes to incubate ~200 µL packed cells in 1 mL buffer B1. 4. Incubate cells with the primary Ab diluted in B1, for 2 h at RT with gentle shaking (see Note 4).

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Adipose Tissue Protocols by Gérard Ailhaud


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