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EZClick™ Global Protein Synthesis Assay Kit (FACS/Microscopy), Green Fluorescence (K459)

April 2018

EZClick™ Global Protein Synthesis Assay Kit (FACS/Microscopy), Green Fluorescence (K459)


MILPITAS, Calif. - April 18, 2018 - PRLog -- Cells generate a complete set of proteins during division. Protein synthesis is a tightly regulated process and many critical controls in gene expression occur at the level of translation to ensure that production of specific cellular proteins is quickly turned on/off under specific conditions (heat shock, starvation, etc.). Protein synthesis is essential in cell growth, proliferation, signaling, differentiation or death; therefore, the identity and amount of the synthesized proteins are critical parameters in determining the physiological state of the cell. Methods enabling detection and characterization of nascent proteins, or changes in spatial and temporal protein expression/degradation patterns during disease, drug treatments or environmental changes are important tools in assessment of cytotoxicity. Biovision's EZClick™ Global Protein Synthesis Assay Kit utilizes a novel and robust chemical method based on an alkyne containing and cell-permeable analog of puromycin, O-Propargyl-puromycin (OP-puro). Once inside the cell, OP-puro stops translation by forming covalent conjugates with nascent polypeptide chains. Truncated polypeptides are rapidly turned over by the proteasome and can be detected based on a click reaction with the fluorescent azide. Unlike methionine analogs, OP-puro does not require methionine-free conditions and can be used to label nascent proteins directly in the cell culture. Our kit provides sufficient materials for 100 assays to detect nascent proteins synthesized under various physiological conditions, and Cycloheximide, an inhibitor of protein synthesis that serves as an experimental control.

Figure: Inhibitory effect of Cycloheximide on nascent polypeptides synthesis. HeLa (105 cells/ ml) and Jurkat (1X106 cells/ml) cells respectively were pre-treated with vehicle or Cycloheximide for 30 min at 37°C. Subsequently, cells were labeled with BioVision's K459. Cells were then processed and analyzed by Microscopy and FACS according to the kit protocol. Green fluorescence (upper panel) corresponds to de novo synthesized polypeptides whereas bottom panel shows the inhibitory effect of Cycloheximide on protein biosynthesis. Nuclear staining in both panels confirms that green signal is a result of Protein Label incorporation.

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About BioVision:

BioVision, Inc. is a privately held Life Science company headquartered in the beautiful San Francisco Bay area. BioVision develops and offers a wide variety of products including assay kits, antibodies, recombinant proteins & enzymes, and other innovative research tools for studying Apoptosis, Metabolism, Cell Proliferation, Cellular Stress, Cell Damage and Repair, Diabetes, Obesity and Metabolic Syndrome, Stem Cell Biology, Gene Regulation, Signal Transduction, and more.

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