Immunofluorescent analysis of Red firefly luciferase (from the Japanese firefly Luciola cruciata) in pCMV-Red Firefly (RFF) Luc transfected (green, left panel) and untransfected (UT) 293 cells (right panel). 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 RFF luciferase polyclonal antibody (Product # PA1-179) 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||Luciferase|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified full length recombinant Red Firefly luciferase expressed in E. coli.|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|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 PA1-179 detects a prominent ~62 kDa protein in Red Firefly luciferase expressing cells. No signal was detected in Green Renilla transfected or untransfected control HEK293 cells. This antibody has also been successfully tested in IP, Immunofluorescence and neutralization assays.
Red Firefly luciferase activity can be measured using the Thermo Fisher Firefly Luciferase Flash
or Glow Assay Kits or Luciferase Dual Assay Kits.
Firefly luciferase is a 62kDa protein produced in nature by several species of the Lampyridae family of beetles, which includes the genera Photinus and Luciola. Thermo Scientific Red Firefly Luciferase has an emission range of 560-700nm (lambda max = 613nm). Bioluminescent signal from firefly luciferase originates from the oxidation of D-Luciferin. Light output captured using a luminometer can be correlated with the amount of firefly luciferase protein produced and used to determine the activity of the promoter driving firefly luciferase expression. Firefly luciferase bioluminescence provides researchers with a non-radioactive assay for measuring transcriptional activity of regulatory elements. Firefly luciferase 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.