Proteasome Sensor Vector

Description

The Proteasome Sensor Vector pZsProSensor-1 encodes the gene forZoanthussp. Green Fluorescent Protein ZsGreen fused to the mouse ornithine decarboxylase MODC degradation domain amino acids 410461 . This vector is designed for studies of proteasome function in mammalian cells: Since the MODC degradation domain targets the constitutively-expressed protein for rapid degradation, the protein does not accumulate in cells until the proteasome is inhibited, which is indicated by an increase in green fluorescence. This vector also contains a kanamycin/neomycin selectable marker, as well as SV40 and pUC origins of replication. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Measuring proteasome activity in living cells.HEK 293 cells stably transfected with the Proteasome Sensor Vector Panel A were selected in G418 and sorted by flow cytometry, and then treated with 10 muM ALLN for the indicated times. ALLN is a peptide aldehyde Ac-Leu-Leu-Nle-al that reversibly inhibits the proteasomes chymotrypsin activity, preventing it from attacking the ZsProSensor-1 protein. As a result, ZsProSensor-1 quickly accumulates, resulting in a strong green emission signal that can be measured by fluorescence microscopy Panel B , flow cytometry Panel C , or fluorometry with a 96-well plate reader Panel D . Back High throughput screening HTS assay to identify proteasome inhibitors.Composite images of the HEK 293 ZsGreen Proteasome Sensor Cell Line were acquired using the ArrayScan HCS reader. The cells were stained with 10 muM Hoechst 33342 blue .Panel A.The positive control bortezomib.Panel B.Disulfiram.Panel C.PDTC.Panel D.Negative control PBS .