BIO 102 Lecture 3 Notes

Bleac h frap wit h las er beam watc h t ime for rec

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Unformatted text preview: hot obleac hing Photobleac hing: bleac h FRAP wit h las er beam, watc h t ime for rec ov ery (bec aus e ev ery thing els e dif fus es ) [ c an hav e rapid rec ov ery or no rec ov ery at all] Early objec tion: put t ing so much energy t hat it behaves dif ferent ly; might cause permanent damage in bleac hed region (introduc tion of FLIP) FLIP! Fluoresc enc e los s in phot obleaching Modes of res t ric t ing lat eral mobilit y -lateral c lus t ering of IMPs -ECM binding/ c lus t ering of IMPs -c y t os k elet al res t raint of IMPs -c ell adhes ion mediated c lus t ering of rec eptors Tet hering is important Ex perimental c harac teriz at ion of int egral and peripheral membrane prot eins : 1) ex trac t peripheral proteins wit h c ombination of t reat ment s t hat dif f erent ially s olubiliz e dif f erent c las s es of peripheral prot eins 2. ex trac t ed membranes (enric hed in int egral membrane prot eins are t reat ed with mild nonionic detergent t o s olubiliz e membrane prot eins , whic h c an t hen be purified by met hods t o be dis c us s ed lat er...
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