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The Cignal Lenti Reporters are ready for transduction right out of the box.
There is no need to generate or propagate lentivirus in your laboratory. These
vectors are extremely useful for transient transduction studies in difficult to
transfect cells or for pathway sensor cell line generation.
Each Lenti Reporter Array includes 10 Cignal Lenti Pathway Reporters and two
controls. All reporters and controls are delivered as transduction-ready
lentiviral particles (250 µl at 1X107 transducing units/ml). All reporter
assays utilize firefly luciferase reporter gene technology.
Firefly luciferase results are calculated for each transduced culture. The
change in the activity of each signaling pathway is determined by comparing the
normalized luciferase activities of the reporter in treated versus untreated
cells.
The identically treated negative control expresses firefly luciferase under
the control of a minimal promoter and serves as a pathway specificity control.
The positive control is a valuable reagent for optimizing your transduction
conditions
Transient Pathway Regulation Studies in Difficult to Transfect Cell Lines:
Target cells are transduced with the Cignal Lenti Pathway Reporter. The cells
are typically cultured for 24 to 48 hours to insure lentivirus integration. The
cultures are then treated with the biological agents of interest (siRNA, shRNA,
chemical compound, viral expression vector, protein, peptide). Reporter assays
are carried out 18 to 36 hours post-treatment,
depending upon the treatment conditions.
Pathway Sensor Cell Line Generation: Target cells are transduced with
the Cignal Lenti Pathway Reporter. Following transduction, the cells are
cultured under puromycin selection to generate a homogenous population of
transduced cells. If necessary, single cell cloning may be carried out in order
to isolate a clonal pathway sensor cell line. These pathway sensor cell lines
serve as a valuable cell-based assay platform, for subsequent screening and
mechanism of action studies.
Simple Procedure:
- Transduce Cignal Lenti Reporters into target cells
- Treat with nucleic acid, protein, peptide, or small molecule of
interest
- Perform reporter quantitation using firefly luciferase activity
assay
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