| Description |
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The Development Reporter Array is the first commercial reporter array that
allows you a comprehensive view of critical growth and differentiation
signaling pathways. Don't rely on guesswork to predict gene or drug
function. Use the Cignal Finder Development Array to pinpoint the pathways
regulated by the genes or drugs your laboratory is studying.
Learn how the Development Reporter Array works
The Cignal Finder Development 10-Pathway Reporter Array is ideally suited
for discovering:
- siRNA/shRNA/miRNA phenotypes
- Biological responses to chemical compounds
- Mechanisms of action of proteins, peptides, and ligands
For more information about the Cignal Finder 10-Pathway Reporter Arrays,
please visit the Cignal Finder Array
home page.
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| Pathway Listing and Performance Data |
| Tube/Column |
Pathway |
Transcription Factor |
| 1 |
Notch |
RBP-Jκ |
| 2 |
Wnt |
TCF/LEF |
| 3 |
Myc/Max |
Myc/Max |
| 4 |
NFκB |
NFκB |
| 5 |
TGFβ |
SMAD2/SMAD3/SMAD4 |
| 6 |
Cell Cycle/pRb-E2F |
E2F/DP1 |
| 7 |
C/EBP |
C/EBP |
| 8 |
cAMP/PKA |
CREB |
| 9 |
MAPK/ERK |
Elk-1/SRF |
| 10 |
MAPK/JNK |
AP-1 |
| 11 |
Cignal negative control |
|
| 12 |
Cignal positive control |
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Learn more about the 10 pathways
View each reporter's transcriptional
response element sequence
Cellular differentiation involves the development of a totipotent or
pluripotent stem cell into a terminally differentiated cell having a
specialized function. This process involves changes to the shape, size,
location, and metabolic activity of the cell. Successful cellular growth and
differentiation depends on a carefully controlled pattern of gene expression
events. The Cignal Finder Development Reporter Array consists
of 10 Cignal Reporter Assays that are used to measure the activities of 10
pathways that function as key regulators of cellular growth and
differentiation.
The Notch signaling pathway plays a critical role in mediating
cell-cell interactions, particularly those important in embryogenesis and
organogenesis. The Wnt family of proteins are potent morphogenic
ligands, and operate in concert with TGFβ, Notch, Hedgehog,
and other signaling pathways. These pathway interactions ultimately activate
the expression of key downstream genes that control apoptosis,
proliferation, morphogenic changes, extracellular matrix remodeling, cell
adhesion and migration. The pRb/E2F pathway serves as a critical cell cycle
checkpoint system, and is the target of numerous mitogenic and
differentiation signals. The MAPK/ERK and MAPK/JNK signaling activities are
precisely tuned during cellular development, in response to various growth,
survival, and differentiation factors. Signaling through these pathways
leads to the regulation of the Myc, NFκB, CREB, AP-1, ELK-1,
SRF and C/EBP
proteins. The activities of each of these transcription factors can be
analyzed using the Cignal Finder Development Reporter Array. The cAMP/PKA
signaling pathway intersects with numerous signal transduction pathways
during cellular development, including the NFκB, PKC/Ca++, and
TGFβ
pathways.
Performance Data
Identify key developmental signaling modulated by your gene of
interest
Notch signaling plays a key role in cell fate determination and
differentiation and its biological function depends on context-specific
interactions with other signaling pathways. Therefore, it is important to
identify the effect of overexpression of the Notch gene on the other
developmentally important signaling pathways in your cell line of choice.
The Cignal Finder Development 10-Pathway Reporter Array is a central tool
for this research.
Development 10-Pathway Reporter Array showed that overexpression of
constitutively active Notch1 specifically up-regulates only the Notch
signaling pathway and has no effect on the other important developmental
signaling pathways in HEK-293H cells.
HEK-293H cells were reverse transfected with the Cignal Finder Development
10-Pathway Reporter Array. After 16 hours of transfection, medium was changed to
assay medium (Opti-MEM containing 0.5% of fetal bovine serum, 1% NEAA, 100 U/ml
Penicillin and 100 µg/ml Streptomycin). After 24 hours of transfection,
cells were infected with 10 MOI of recombinant adenoviruses expressing
constitutive active Notch1 (Ad-NICD) or 10 MOI of recombinant adenovirus
expressing GFP (Ad-GFP). Dual-Luciferase assays was performed 18 hours after
infection, and results are expressed as fold change. The fold change was
calculated by dividing the normalized luciferase activities of each
pathway-focused reporter infected with the Ad-NICD virus by the normalized
luciferase activity of the respective pathway-focused reporter infected with the
Ad-GFP virus. Experiments were done in quadruplicates, and the standard
deviations are indicated.
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| How It Works
|
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Each Reporter Array includes 10 Cignal Reporter Assays and two controls in
either tube or plate format. All reporter assays are based on dual-luciferase
technology. Each reporter consists of a mixture of a pathway-focused
transcription factor-responsive firefly luciferase construct and a
constitutively expressing Renilla luciferase construct.
Dual-luciferase results are calculated for each transfectant. The change in
the activity of each signaling pathway is determined by comparing the normalized
luciferase activities of the reporter in treated versus untreated transfectants.
The identically treated negative control serves as a specificity control. The
positive control serves as a control for transfection efficiency, by monitoring
GFP expression, as well as a positive control for both the firefly and Renilla
luciferase assays.
Simple Procedure:
- Transfect Cignal Reporter Assays and test nucleic acids into cells
- Treat with protein, peptide, or small molecule of interest
- Perform reporter quantitation using luciferase activity assays
Cignal Finder Array - Tube Format
The tube format of the Cignal Finder Arrays are delivered in 12-tube
strips, along with important negative and positive controls. The assays
are used right out of the box for the transfection or reverse
transfection of the reporter assays into your cell lines of interest.
Cignal Finder Array - Plate Format
The plate format of the Cignal Finder Arrays are delivered in a 96-well
cell culture plate. Each reporter and control assay is dried down in
each column of the plate (8 wells per assay).
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| Useful Links |
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| Related Products |
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| Brief Protocol: For Experienced
Users |
First time users, please refer to the complete
protocol in the Cignal Reporter Assays User Manual.
- Dilute transfection-ready reporter, negative control, and positive
control construct formulations.
- Dilute relevant test nucleic acids (siRNA, shRNA, miRNA, expression
vector).
- Prepare appropriate combinations of reporter constructs, controls,
and test nucleic acids.
- Transfect plasmid mixtures separately into replicate wells of your
cell line of interest using an optimized transfection procedure for
the cell line under study.
- If applicable, 16 to 24 hours post-transfection, treat the
transfected cells with test proteins, peptides, or compounds of
interest.
- Two (2) to three (3) days post-transfection, assay the activities of
the signaling pathways under study, utilizing the dual luciferase
assay.
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