Iodixanol Gradient Centrifugation to Separate Components of the Low-Density Membrane Fraction from 3T3-L1 Adipocytes.
| Autor: | Sadler JB; University of Glasgow, Glasgow G12 8QQ, United Kingdom;, Lamb CA; Cancer Research UK, London Research Institute, London EC1V 4AD, United Kingdom;, Gould GW; University of Glasgow, Glasgow G12 8QQ, United Kingdom;, Bryant NJ; University of York, Heslington, York YO10 5DD, United Kingdom. |
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| Jazyk: | angličtina |
| Zdroj: | Cold Spring Harbor protocols [Cold Spring Harb Protoc] 2016 Feb 01; Vol. 2016 (2), pp. pdb.prot083709. Date of Electronic Publication: 2016 Feb 01. |
| DOI: | 10.1101/pdb.prot083709 |
| Abstrakt: | We optimized a set of fractionation techniques to facilitate the isolation of subcellular compartments containing insulin-sensitive glucose transporter isoform 4 (GLUT4), which is mobilized from GLUT4 storage vesicles (GSVs) in fat and muscle cells in response to insulin. In the absence of insulin, GLUT4 undergoes a continuous cycle of GSV formation and fusion with other compartments. Full membrane fractionation of 3T3-L1 adipocytes produces a low-density membrane fraction that contains both the constitutive recycling pool (the endosomal recycling compartments) and the insulin-sensitive pool (the GSVs). These two pools can be separated based on density using iodixanol gradient centrifugation, described here. (© 2016 Cold Spring Harbor Laboratory Press.) |
| Databáze: | MEDLINE |
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