Abstract
Metabolic labeling of mucins is a powerful method for two reasons: (1) it lowers the detection limits of the mucins and their precursors considerably, and (2) it provides data on the actual synthesis of mucins in living cells. The produced radioactive mucins can be isolated and studied using biochemical methods, as described in Chapter 19, but these techniques apply basically to the study of mature mucins. In this chapter, we outline the methods for the immunoprecipitation of mucins, i.e., the immunoisolation of the mature mucins as well as their corresponding precursors. By applying metabolic labeling using amino acids and immunoprecipitation with the proper antibodies against the mucin polypeptide, it becomes possible to detect the earliest mucin precursor in the rough endoplasmic reticulum, to follow its subsequent complex conversion into a mature mucin, and to observe its storage and eventual secretion (1–3). Moreover, this antibody-based technique has the required specificity to discriminate the primary translation-product of each mucin gene. How mucin precursors can be distinguished is described in detail for each of the MUC-type mucins in Chapter 21 .
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Van Jan-Willem Klinken, B., Büller, H.A., Einerhand, A.W.C., Dekker, J. (2000). Identification of Mucins Using Metabolic Labeling, Immunoprecipitation, and Gel Electrophoresis. In: Corfield, A.P. (eds) Glycoprotein Methods and Protocols. Methods in Molecular Biology™, vol 125. Humana Press. https://doi.org/10.1385/1-59259-048-9:239
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DOI: https://doi.org/10.1385/1-59259-048-9:239
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