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. Intended Use:Detection of nascent protein synthesisDetection of protein expression and degradation patterns in presence of cytotoxic agents Screening for genotoxic compounds and effectors of protein synthesisSample Type:Suspension or adherent cell culturesSpecificity:MammalianTest Principle:The Protein Synthesis Monitoring BioAssay 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. This kit provides sufficient materials for 50 assays to detect nascent proteins synthesized under various physiological conditions,and Cycloheximide,an inhibitor of protein synthesis that serves as an experimental control.Kit Components:Wash Buffer (10X),1x25mlFixative Solution,1x10mlPermeabilization Buffer (10X),1x25mlProtein Label (400X),1x125ulCopper Reagent (500X),1x100ulFluorescent Azide (100X),1x250ulReducing Agent (20X),1x1.25mlCycloheximide (100X),1x100ulStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product,centrifuge the original vial after thawing and prior to removing the cap.