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Immunofluorescent analysis of Green Renilla luciferase (from the Sea pansy Renilla reniformis) in untransfected (UT, left panel) and pCMV-Green Renilla (GR) luciferase transfected (green, right panel) 293 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 2% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a GR luciferase polyclonal antibody (Product # PA1-180) at a dilution of 1:1000 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Renilla|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified full length recombinant Green Renilla luciferase expressed in E. coli.|
|Storage buffer||PBS with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:500 - 1:1000|
|Immunoprecipitation (IP)||5 µg|
|Neutralization (Neu)||Assay Dependent|
|Western Blot (WB)||1:2000 - 1:4000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Western blot analysis of PA1-180 detects a prominent ~36 kDa protein in Green Renilla luciferase expressing cells. Additional unspecific bands are also detected in control cells with increased exposure time. This antibody has also been successfully tested in IP, Immunofluorescence, and neutralization assays.
Green Renilla luciferase activity can be measured using the Thermo Fisher Firefly Luciferase Flash or Glow Assay Kits or Luciferase Dual Assay Kits.
The Green Renilla luciferase is a 36kDa protein produced by a derivative of the wild type Renilla luciferase gene from the sea pansy, Renilla reniformis. Compared to the wild type luciferase, Green Renilla is more stable in serum and has an the emission spectrum that is shifted toward the green region. The protein provides extremely bright flash signal that decays rapidly. The Renilla Flash and Glow Assay Kit reagents allow for Glow (stabilized signal with ~3 hours halflife) or Flash type bioluminescence experiments and are optimized for best results with Thermo Scientific Green Renilla Luc Plasmids. Bioluminescence provides researchers with a non-radioactive assay for measuring transcriptional activity of regulatory elements. Reporter activity can be used to determine promoter activity, to study upstream pathway activity in the cell, or to determine effect of activators or inhibitors.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
EC 188.8.131.52; Green-shifted Renilla luciferase; Renilla-luciferin 2-monooxygenase; Renilla-type luciferase