Science Catalog 2012 Life Cell Signaling Reporter Vectors and Cell Lines pGL4 Firefly Luciferase Reporter Vectors Selection Guide • Contains luc2 gene (Photinus pyralis): Codon optimized for mammalian expression. Features >90% fewer cryptic sites than luc+ (pGL3 Vectors). • Updated Vector backbone with 78% fewer cryptic sites than pGL3. • Improved MCR with additional RE sites. • Available in Rapid Response™ Reporter configurations. • See the online pGL4 Vector Selector at: faqs.promega.com and choose the Solution Finder tab. YES (time and fold induction) preferred, even with lower absolute RLUs? Is maximum responsiveness YES pGL4.12[luc2CP] Vector NO Do you require a reporter that displays higher fold changes in less time, even at the expense of lower absolute expression? YES pGL4.11[luc2P] Vector NO pGL4.10[luc2] Vector [luc2/SV40] Vector pGL4.13 pGL4.10[luc2] Vector [luc2/SV40] Vector pGL4.13 For a positive control/reference. Will you use the luc2 gene as an experimental reporter in which to clone regulatory elements of interest? NO For a negative control/baseline reference. For a transfection control to Renilla. pGL4 Renilla Luciferase Reporter Vectors Selection Guide • Contains hRluc gene (Renilla reniformis). • Updated Vector backbone with 78% fewer cryptic sites than pGL3/phRG. • Available in Rapid Response™ Reporter configurations. • See the online pGL4 Vector Selector at: faqs.promega.com and choose the Solution Finder tab. YES Considerations for Choosing a Control • Relative Light Units of the co-reporter generally should be at the lowest level necessary to quantitate reliably. Generally speaking, the order of vector expression is pGL4.72[hRlucCP] <pGL4.71[hRlucP] <pGL4.70[hRluc] <pGL4.74[hRluc/TK] <pGL4.73[hRluc/SV40] <pGL4.75[hRluc/CMV]. The order and level of expression can depend upon cell type and other factors. • Promoter cross-talk interference. A strong promoter may cause a so-called “squelching” effect— soaking up general transcription factors—or it may specifically interfere with the test promoter by competing for the same factor(s) that is (are) required for transcription for both promoters. SV40 and CMV are generally strong promoters and thus may not be suitable for use as control reporters. • Undesirable regulation of co-reporter expression by experimental stimulus. Testing different vector configurations is recommended to minimize the impact of undesirable regulation of the control reporter. Is low to moderate constitutive expression appropriate for your application? YES NO Is strong, constitutive expression appropriate as a control? YES NO Is baseline expression appropriate for your application? YES NO Do you require a reporter that displays higher fold changes in less time, even at the expense of lower absolute expression? YES Is maximum responsiveness (time and fold induction) preferred, even with lower absolute RLUs? YES pGL4.74[hRluc/TK] Vector pGL4.75[hRluc/CMV] Vector pGL4.73[hRluc/SV40] Vector pGL4.70[hRluc] Vector pGL4.72[hRlucCP] Vector NO pGL4.71[hRlucP] Vector For negative control/baseline reference. For positive control/ reference; is maximum constitutive expression appropriate? YES pGL4.70[hRluc] Vector pGL4.75[hRluc/CMV] Vector NO pGL4.73[hRluc/SV40] Vector NO Are you studying a full promoter or regulatory element? YES NO Will you use the pGL4 Renilla Vector as a co-reporter to normalize transfection efficiency, cytotoxicity or other variables? NO 270 For complete and up-to-date product information visit: www.promega.com/catalog 4312MA09_3B Pagina 273
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