Download Molecular Motors: Methods and Protocols (Methods in by Ann O. Sperry PDF

By Ann O. Sperry

Molecular motor proteins produce strength for circulate in a really big range of mobile procedures. This quantity explores the intense practical and structural range of molecular automobiles and offers tools correct to every motor kin. moreover, it describes innovations directed at automobiles that fall outdoors of the 3 characterised households: dynamin and F1ATPase.

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Additional resources for Molecular Motors: Methods and Protocols (Methods in Molecular Biology Vol 392)

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Disconnect the power supply, remove the upper buffer chamber with the attached gels, and discard the buffer. 13. Remove the gel sandwich from the casting stand and lay it on a paper towel. Carefully pry open the glass plates, and cut a small triangle at one corner of the gel to mark the orientation of the gel. 14. To visualize the proteins, immerse the gel in Coomassie blue staining solution for 1 h with gentle shaking, and then briefly rinse the gel with deionized water and transfer it to destaining solution until the gel is completely destained.

Detergent stocks: Triton X-100 stocks are prepared as 10% w/v. Long-term storage of detergent solutions is not recommended as this can lead to formation of peroxide radicals. 1. Metabolic Labeling Experiments 1. 5% Triton X-100, 80 mM β-glycerophosphate, 50 mM NaF, and 100 mM potassium phosphate. 2. Complete lysis buffer (CLB): Prepare immediately before experiments by adjusting the volume of 1X LB buffer needed to 2 mM sodium orthovanadate, 100 nM staurosporine, 100 nM K252a, 50 nM okadaic acid, 50 nM microcystin, 1/100 dilution of PIC, and 1/100 dilution of mammalian protease inhibitor cocktail (see Note 2).

Biol. 17, 82–88. , 5. , and Wordeman, L. (2004) A standardized kinesin nomenclature. J. Cell Biol. 167, 19–22. A. 6. (2005) Kar3 interaction with Cik1 alters motor structure and function. EMBO J. 24, 3214–3223. P. 7. (2005) Cik1 targets the minus-end kinesin depolymerase kar3 to microtubule plus ends. Curr. Biol. 15, 1420–1427. E. (1999) Kin I kinesins 8. are microtubule-destabilizing enzymes. Cell 96, 69–78. 9 Wordeman, L. J. (1995) Identification and partial characteriza9. tion of mitotic centromere-associated kinesin, a kinesin-related protein that associates with centromeres during mitosis.

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