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Improving structural integrity of cryosections for immunogold labeling
It is found that the step during which sections are thawed and transferred from the cryochamber to the supporting film on an EM grid is most critical for structural preservation, and substituting a mixture of methylcellulose and sucrose for the conventional sucrose transfer medium was able to alleviate the problem of overstretching, thus improving greatly the structural integrity of thin cryosections. Expand
Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat
The results suggest that in the presence of insulin GLUT 4 recycles from the cell surface, probably via the coated pit-endosome pathway that has been characterized for cell surface receptors, and also that insulin causes the redistribution ofGLUT 4 by stimulating exocytosis from GLUT 2-containing tubulo-vesicular structures, rather than by slowing endocytotic of GLUT 3. Expand
Proteomic and Biochemical Analyses of Human B Cell-derived Exosomes
Data suggest that recruitment of membrane proteins from the limiting membranes into the internal vesicles of multivesicular bodies may involve their incorporation into tetraspanin-containing detergent-resistant membrane domains. Expand
Cryosectioning and immunolabeling
Cryoimmunolabeling methods for the subcellular localization of proteins and certain lipids and the detailed procedures that have been developed and tested in practice in the laboratory during the past decades are described. Expand
A new method of preparing gold probes for multiple-labeling cytochemistry.
  • J. Slot, H. Geuze
  • Biology, Medicine
  • European journal of cell biology
  • 1 July 1985
The usefulness of the new gold probes bound to protein A for multiple labeling in a current immunocytochemical study on receptor mediated transport of IgA in human duodenal crypt cells is demonstrated. Expand
Vesicular Tubular Clusters between the ER and Golgi Mediate Concentration of Soluble Secretory Proteins by Exclusion from COPI-Coated Vesicles
Data suggest an important role of VTCs in soluble cargo concentration by exclusion from COPI-coated domains, as well as KDELr and rBet1, which showed a first, significant concentration in COPII- coated ER buds and vesicles and were prominently present in COPI -coated tips of V TC tubules. Expand
Recycling Compartments and the Internal Vesicles of Multivesicular Bodies Harbor Most of the Cholesterol Found in the Endocytic Pathway
Cholesterol displays a highly differential distribution in the various membrane domains of the endocytic pathway of human B lymphocytes, which is almost absent from lysosomes and present in two types of multivesicular bodies. Expand
Translocation of the glucose transporter GLUT4 in cardiac myocytes of the rat.
Increased glucose transport in muscle and fat is accounted for by translocation of GLUT4 from the intracellular tubulo-vesicular elements to the plasma membrane by stimulation of the exocytotic rate constant. Expand
Intracellular site of asialoglycoprotein receptor-ligand uncoupling: Double-label immunoelectron microscopy during receptor-mediated endocytosis
The tubular vesicles are, it is proposed, an intermediate in recycling the receptor to the cell surface in rats infused with asialoglycoprotein for 60 min, and appear to transform into secondary lysosomes, wherein the ligand is degraded. Expand
Cytotoxic T lymphocyte granules are secretory lysosomes, containing both perforin and granzymes
The subcellular localization of perforin, granzymes, and known endosomal and lysosomal marker proteins was determined in human and murine CTL, by immunogold labeling of ultrathin cryosections followed by electron microscopy, and these observations define CTL granules as secretory lysOSomes. Expand