Fucose (FucAz) Modified Glycoprotein Assay Kit (ab241020)
Key features and details
- Detection method: Fluorescent
- Platform: Flow cytometer, Fluorescence microscope
- Sample type: Adherent cells, Suspension cells
Overview
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Product name
Fucose (FucAz) Modified Glycoprotein Assay Kit -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells -
Product overview
Fucose (FucAz) Modified Glycoprotein Assay Kit (ab241020) is a highly specific, simple and robust method for labeling and detection of fucosylated proteins within cells. It uses a fucose analog that is fed directly into the cells, processed via the fucose salvage pathway and incorporated into the glycoproteins. This is followed by click reaction with an alkyne-containing dye so 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. The kit provides sufficient materials for 100 assays in a 96-well plate format.
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Platform
Flow cytometer, Fluorescence microscope
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Copper Reagent (100X) 1 x 100µl Fixative Solution 1 x 10ml Fluorescent Alkyne (100X) 1 x 100µl FucAz Label (1000X) 1 x 10µl Permeabilization Buffer (10X) 1 x 25ml Reducing Agent (20X) 1 x 500µl Total DNA Stain (1000X) 1 x 20µl Wash Buffer (10X) 1 x 25ml -
Relevance
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.
Images
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Analysis of metabolic labeling of FucAz labeled glycans in proliferating cells
Jurkat cells (1X106 cells/ml) were cultured in presence of 1X 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).
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Analysis of metabolic labeling of FucAz labeled glycans in proliferating cellsFluorescence Microscope images of subcellular localization (middle panel) of Fucosylated glycoproteins in fixed HeLa cells. Nuclear staining (left panel) and composite image (right panel).