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By Weiming Xia, Huaxi Xu

ISBN-10: 0849322456

ISBN-13: 9780849322457

Within the look for a good therapy for Alzheimer's sickness, APP is a special version protein that illustrates the big choice of easy and complex characterization strategies on hand.

Exploring numerous organic ideas to explain the constitution and serve as of this transmembrane protein, this article offers every one procedure with distinctive, step by step protocols to accomplish reproducible effects and supply a framework for learning different membrane proteins.

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Fm Page 19 Wednesday, November 3, 2004 2:59 PM 2 Assays for Analysis of APP Secretion and Recycling Markus P. Kummer, Tsuneo Yamazaki, and Edward H. 4 Discussion Acknowledgments References ABSTRACT The trafficking of the amyloid precursor protein (APP) involves the concomitant secretion and endocytosis of APP from the cell surface. In addition, APP recycles between the endocytic compartment and the cell surface. This complex sequence of events can be studied by a reliable and reproducible assay based on the binding of a radiolabeled APP antibody.

A 5-ml quantity of GS115 is grown overnight in YPD in a 50-ml conical flask at 30oC. 5. On the following day, inoculate the overnight culture into 500 ml YPD in a 2-liter flask. 5. 6. Centrifuge (1500 × g for 5 min) and resuspend cells in 500 ml ice cold water. 7. Centrifuge and resuspend in 250 ml ice-cold water. 8. Centrifuge and resuspend in 20 ml ice-cold 1 M sorbitol. 9. Centrifuge and resuspend in 1 ml ice-cold 1 M sorbitol. 10. 2-cm electroporation cuvette on ice. Electroporate according to manufacturer’s recommended conditions for yeast.

Chem. 268, 608, 1993. 3. J. Trafficking of cell-surface amyloid betaprotein precursor. II. Endocytosis, recycling and lysosomal targeting detected by immunolocalization. J. Cell Sci. 109, 999, 1996. 4. Simons, M. et al. Cholesterol depletion inhibits the generation of beta-amyloid in hippocampal neurons. Proc. Natl. Acad. Sci. USA 95, 6460, 1998. 5. E. et al. Effect of increased glycogen synthase kinase-3 activity upon the maturation of the amyloid precursor protein in transfected cells. Neuroreport 8, 639, 1997.

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Amyloid precursor protein : a practical approach by Weiming Xia, Huaxi Xu

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