Science Catalog 2012 Life Cell Signaling psiCHECK™-1 and psiCHECK™-2 Vectors Product psiCHECK™-1 Vector psiCHECK™-2 Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The psiCHECK™-1 and psiCHECK™-2 Vectors are designed to provide a quantitative and rapid approach for initial optimization of RNA interference (RNAi). The vectors enable monitoring of changes in expression of a target gene fused to a reporter gene. In both vectors Renilla luciferase is used as the primary reporter gene, and the gene of interest is cloned into a multiple cloning region located downstream of the Renilla translational stop codon. Initiation of the RNAi process by synthetic siRNAs or in vivo-expressed shRNAs toward a gene of interest results in cleavage and subsequent degradation of the fusion mRNA. Measuring decreases in Renilla activity provides a convenient way of monitoring the RNAi effect. In comparison with other fusion approaches (e.g., GFP or flag-tags), the Renilla luciferase approach offers more convenient and rapid quantitation with higher sensitivity. The psiCHECK™-1 Vector is recommended for use in monitoring RNAi effects in live cells. The changes in Renilla luciferase activity are measured with the EnduRen™ Live Cell Substrate (Cat.# E6481), which allows continuous monitoring of intracellular Renilla luminescence. The psiCHECK™-2 Vector contains a second reporter gene, firefly luciferase, and is designed for endpoint lytic assays. Introduction of firefly luciferase in the psiCHECK™-2 Vector allows normalization of Renilla luciferase expression, achieving robust and reproducible results. Features: • Save Money: Quantitation is performed with a common luminometer; no need to purchase expensive equipment. • Choose Your Format: Protocols allow for measurements in live cells or crude cell lysates. • Save Time: No requirement for labor-intensive, time-consuming assays or waiting for phenotypic changes. • Convenient: No requirement for transfection normalization when using the psiCHECK™-2 Vector. Storage Conditions: Store at –20°C. Protocol siCHECK™ Vectors Part# TB329 1674 NotI BamHI 1738 1663 PmeI 1654 EcoRI 1640 SgfI 1643 XhoI 1651 SmaI hRluc Promoter T7 NheI 684 BamHI 4451 SV40 Late poly(A) Ampr hluc+ ori HSV-TK promoter 1640 SgfI 1643 XhoI 1663 PmeI 1674 NotI Synthetic poly(A) psiCHECK™-2 Vector (6273bp) hRluc Promoter T7 NheI 684 BglII 1 KpnI 58 SV40 early enhancer/ promoter Ampr ori Synthetic poly(A) psiCHECK™-1 Vector (3560bp) BglII 1 KpnI 58 SV40 early enhancer/ promoter HindIII 420 PstI 462 5´ + 5´ translation 3´ 5´ cleavage of mRNA degradation hRluc light hRluc no light Mechanism of action of the psiCHECK™ Vectors. 3´ mRNA shRNA (21–23bp) 3´ Size Cat.# Price (Fr) 20 µg C8011 538.00 20 µg C8021 581.00 hRluc translation stop gene of interest 292 For complete and up-to-date product information visit: www.promega.com/catalog 4345MA10_3A 4339MA10_3A 4343MA10_3A Pagina 295
Pagina 297Voor artikelen, online handleidingen en verenigingsbladen zie het Online Touch content management system systeem. Met de mogelijkheid voor een online shop in uw spaarprogramma.
Promega Switzerland - Life Sciences Catalog 2012 Lees publicatie 100Home