Product Details
| abID | ab235634 |
|---|---|
| Cat # +Size | K715-100 |
| Size | 100 assays |
| Kit Summary | • Detection method- Flow Cytometry (Ex/Em 440/(540/580) nm) and Fluorescence Microscopy • Sample type- Adherent and suspension cells • Species reactivity- Mammalian • Applications- This assay provides a convenient and accurate procedure to measure de novo Protein synthesized in biological samples. |
| Detection Method | Flow Cytometry (Ex/Em (440/490)/(530/590) nm) and Fluorescence Microscopy |
| Species Reactivity | Eukaryotes |
| Applications | This assay provides a convenient and accurate procedure to measure de novo protein synthesized in biological samples. |
| Features & Benefits | Simple, fast, does not require lengthy incubation times |
| Kit Components | • EZClick™ Wash Buffer (10X) • Fixative Solution • Permeabilization Buffer (10X) • EZClickTM Protein Label (400X) • Copper Reagent (100X) • EZClick™ Fluorescent Azide (100X) • Reducing Agent (20X) • EZClick™ Total DNA Stain (1000X) • Cycloheximide (100X) |
| Storage Conditions | -20 °C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
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™ Protein Synthesis Monitoring 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.
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What is the role of cyclohexamide in K715 and Do I need to add Cycloheximide for every test?
Cycloheximide is an inhibitor of protein synthesis. It serves as an experimental control and it is good to have 2 wells of experimental control for the experiment.
Can we add EDTA to the cell media after cyclohexamide treatment?
Unfortunately, EDTA cannot be used in this assay as it is not compatible with the labeling part of the assay.
