EZClick™ Sialic Acid (ManAz) Modified Glycoprotein Assay Kit (FACS/Microscopy, Green Fluorescence)

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. Intracellular glycans mediate protein folding, stability, and trafficking while at the cell surface, they participate in recognition, cell-cell interactions involved in adhesion, migration, and embryonic development; host-pathogen interactions critical for bacterial and viral infections; and initiation of immune response. Aberrant glycosylation profiles correlate with inflammation and are universal feature of cancer, with sialic acids playing an especially prominent role as tumor associated carbohydrate antigens (TACAs). Altered sialylation of tumor cell surfaces is associated with several critical malignant properties that include invasiveness and metastatic potential suggesting its implication in clinical diagnosis. Since glycoproteins are not directly encoded in the genome, methods of characterization and analyses of glycoproteins are of great interest. Thus BioVision offers EZClickTM Sialic Acid (ManAz) Modified Glycoprotein Assay Kit, a highly specific, simple and robust method for labeling and detection of N-linked glycosylation of cell surface proteins. We use a modified mannosamine precursor that is fed directly into the cells, converted to sialic acid by the sialic acid biosynthetic machinery, and transported to the Golgi apparatus for glycan elaboration. 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. 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