Metabolic radiolabeling of animal cell glycoconjugates.
نویسندگان
چکیده
This unit describes metabolic labeling techniques that provide specific information about the structure, sequence, and distribution of the sugar chains of glycoconjugates. Although these techniques provide less information than complete sequencing of the sugar chains, the partial structural information derived is sufficient for many purposes. In the basic procedure presented in this unit, actively growing cell cultures are grown through several population doublings in complete medium supplemented with radiolabeled glycoconjugate precursors to reach a steady-state level of incorporation. In alternate protocols, cells are cultured for a short period of time in a deficient medium that contains a high concentration of radiolabeled precursor. A pulse or pulse-chase labeling procedure can be used to analyze precursor-product relationships. With sequential pulse-labeling, it is possible to obtain quantities of labeled glycoconjugates with the use of a minimal amount of labeled precursor by using the same medium to pulse-label a series of cultures. A support protocol describes the preparation of multiply deficient medium (MDM) for use in making appropriate deficient media.
منابع مشابه
Developmental Changes of the Notochord and its Inductive Effects on the Adjacent Embryonic Germ Layers with Regard to the Role of Glycoconjugates
Notochord is an axial structure derived of embryonic mesoderm and in addition to structural supporting role in inducing nearby germinal layers, it has a basic role in formation of organs such as vertebral column, axial vessels, neural tube and primitive gut. This organ undergoes essential changes during the development process. First, arises from the primitive node and terms notochordal process...
متن کاملCisplatin Induce Changes of Cell Surface Glycoconjugates in Germinal Epithelium of Seminiferous Tubules
Introduction: Cisplatin is a platinum-based drug widely used for the treatment of different cancers. Cell surface glycoconjugates play an important role in cell-cell interactions. The present investigation was carried out to study the toxic effects of double dose injection of cisplatin on cell surface glycoconjugates in rat as an experimental model. Methods: In this experimental study, 45 adul...
متن کاملCorrelation of Retinole Acid-enhanced Sialyltransferase Activity and Glycosylation of Specific Cell Surface Sialoglycoproteins with Growth Inhibition in a Murine Melanoma Cell System1
Retinole acid inhibits the proliferation of the murine melanoma clone S91-C-2 cells, enhances the glycosylation of specific cell surface Sialoglycoproteins, and stimulates Sialyltransferase activ ity. Mutant clones, selected from the S91 -C-2 cells for resistance to the growth-inhibitory effect of retinoic acid, were used to explore whether cell surface modulation by retinoic acid is related to...
متن کاملمطالعه قند انتهایی اِل- فوکوز در گلیکوکونژوگههای سلولی در آدنوکارسینومای کولون
Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 !mso]> st1":*{behavior:url(#ieooui) } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal" mso-tst...
متن کاملAnalysis of cancer-associated colonic mucin by ion-exchange chromatography: evidence for a mucin species of lower molecular charge and weight in cancer.
Cancer-associated mucins in the colon are antigenically distinct and glycosylated differently from their normal counterparts. Mucin-rich glycoconjugate preparations were made from nine non-neoplastic colons, seven colon cancers, and two different xenografts from mucin-producing human colon cancer cell lines, and radiolabeled with 3H. The preparation was applied to a DEAE-cellulose ion-exchange ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Current protocols in protein science
دوره Chapter 12 شماره
صفحات -
تاریخ انتشار 2001