MILPITAS, Calif. -
March 6, 2018 -
PRLog -- Glycans are vital components of glycoproteins, glycolipids, and proteoglycans in all domains of life. Glycosylation occurs co- or post-translationally on >50% of eukaryotic proteins resulting in membrane-associated, intracellular, or secreted glycoproteins that are crucial in cellular processes, protein bioactivity and metabolic turnover. Glycoproteins are grouped by the type of carbohydrate and amino acid linkage site. Fucose is found in glycoproteins and glycolipids present in vertebrates, invertebrates, plants, and bacteria. It is often a terminal sugar in glycans participating in cell–cell signaling and migration implicated in physiological and pathological processes such as fertilization, embryogenesis, lymphocyte trafficking, immune responses, and cancer metastasis. Most target proteins undergoing fucosylation are secretory or membrane proteins on the cell surface. Fucosylation is one of the most important modifications involved in cancer and inflammation. Several types of biomarkers containing fucose are linked to cancer e.g. alpha-fetoprotein (AFP) is used in the diagnosis of hepatocellular carcinoma and surface glycosphingolipid Globo H is an epitope found on the cell surface of breast, prostate, and ovarian cancers. Since glycoproteins are not directly encoded in the genome, methods of their characterization and analyses are of great interest. Thus, BioVision offers EZClick™Fucose (FucAz) Modified Glycoprotein Assay Kit, a highly specific, simple and robust method for labeling and detection of fucosylated proteins within cells. We use fucose analog that is fed directly into the cells, processed via fucose salvage pathway and incorporated into the glycoproteins. Followed by click reaction with alkyne-containing dye, this system offers a powerful method for imaging the localization, trafficking, and dynamics of glycans, or detection by FACS for quantitative studies. Fucose-labeled glycoproteins can be directly detected in 1D or 2D gels using the appropriate excitation sources, or enriched by immunoprecipitation with biotin-alkyne or antibodies prior to proteomic analysis. We provide sufficient materials for 100 assays in a 96-well plate format.
Figure: Analysis of metabolic labeling of FucAz-labeled glycans in proliferating cells: Jurkat cells (1X106 cells/ml) were cultured in presence of 1X EZClick™ FucAz Label for 72 hours at 37°C. Modified glycoproteins were detected according to the kit protocol and green fluorescence was analyzed by FACS (FL-1 channel). Negative control (white line), Background control (green line), Positive control - fluorescence corresponding to fucosylated glycoproteins (pink line), Tunicamycin-
induced suppression of fucosylation (yellow line).
For complete information on this assay kit, visit:
https://www.biovision.com/ezclicktm-fucose-fucaz-modified...
