Fucose (FucAz) Modified Glycoprotein Assay Kit (FACS/Microscopy, Green Fluorescence) (BN00819)

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BN00819
  • Fucose (FucAz) Modified Glycoprotein Assay Kit (FACS/Microscopy, Green Fluorescence) (BN00819)
  • Fucose (FucAz) Modified Glycoprotein Assay Kit (FACS/Microscopy, Green Fluorescence) (BN00819)
€565

Description

ELISA Kit Technical ManualMSDS

Fucose (FucAz) 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. 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, Assay Genie offers 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. A. Jurkat cells (1X10 6 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). B. Fluorescence Microscope images of subcellular localization (middle panel) of Fucosylated glycoproteins in fixed HeLa cells. Nuclear staining (left panel) and composite image (right panel).

Key Information Description

Product SKU

BN00819

Size

100 assays

Kit Summary

  • Detection method- Flow Cytometry (Ex/Em 488/(530/590) nm) and Fluorescence Microscopy
  • Sample type- Adherent and suspension cells
  • Species reactivity- Mammalian
  • Applications- This assay provides a convenient and accurate procedure to measure glycoproteins (Fucosylation) in biological samples.

Detection Method

Flow Cytometry (Ex/Em 480/(530/590) nm) and Fluorescence Microscopy

Species Reactivity

Mammalian

Applications

This assay provides a convenient and accurate procedure to measure glycoproteins (Fucosylation) in biological samples

Features and Benefits

Simple, fast, does not require lengthy incubation times

Kit Components

  • Wash Buffer (10X)
  • Fixative Solution Permeabilization Buffer (10X)
  • GlcAz Label (1000X)
  • Copper Reagent (100X)
  • Fluorescent Alkyne (100X)
  • Reducing Agent (20X)
  • Total DNA Stain (1000X)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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