| Description |
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The Cancer Reporter Array is the first commercial reporter array that allows
you a comprehensive view of cancer-related signaling pathways. Don't rely on
guesswork to predict gene or drug function. Use the Cignal Finder Cancer
Array to pinpoint the pathways regulated by the genes or drugs your
laboratory is studying.
Learn how the Cancer Reporter Array works
The Cignal Finder Cancer 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 |
Wnt |
TCF/LEF |
| 2 |
Notch |
RBP-Jκ |
| 3 |
p53/DNA Damage |
p53 |
| 4 |
TGFβ |
SMAD2/3/4 |
| 5 |
Cell cycle/pRb-E2F |
E2F/DP1 |
| 6 |
NFκB |
NFκB |
| 7 |
Myc/Max |
Myc/Max |
| 8 |
Hypoxia |
HIF1A |
| 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
Cancer is a progressive disease marked by uncontrolled cell growth, invasion
of a tumor mass into neighboring tissues, and metastasis of the
cancerous cells throughout the body via the circulatory and/or lymphatics
systems. Oncogenes and tumor suppressor genes are two classes of genes that
are frequently mutated in cancerous cells. Their gene products serve as
critical regulators of cell proliferation, growth, DNA repair,
vasculogenesis, and cell migration. The 10 Cignal Reporter Assays included
in the Cignal Finder Cancer Reporter Array measure the activities of pathways
that serve as key regulators of oncogenesis and tumor suppression.
These include the well known "guardian of the genome", the
tumor suppressor p53. The MAPK/ERK and MAPK/JNK
signaling pathways are strongly stimulated by environmental and genotoxic
stress, and are key regulators of cellular proliferation, survival,
apoptosis, and tumorigenesis. One of the primary targets of the MAPK
pathways is the Myc transcription factor family. These proteins are
well established proto-oncogenes and control the expression of a wide
variety of genes involved in cellular homeostasis and growth. Another target
of the MAPK signaling machinery is the HIF1α transcription factor, which
activates a number of hypoxia-responsive genes, including those controlling
proliferation and vascularization. The NFκB transcription factors are
constitutively active in many tumors, particularly those of hematopoietic
and neuronal origin. The Rb/E2F pathway is a critical regulator of cell
cycle progression, DNA synthesis, and DNA repair. Notch, Wnt, and TGFβ
function cooperatively to control cellular proliferation, apoptosis,
cell-cell interactions, tumor invasion and metastasis.
Performance Data
Identification of signaling pathway(s) impacted by p53 siRNA treatment
The most heavily studied tumor suppressor gene is p53. To understand more
about the biological function of p53 in the cell line of your choice, it is
important to know the signaling pathways perturbed by knockdown of p53. The
Cignal Finder Cancer 10-Pathway Reporter Array provides a vital tool to
identify the key cancer signaling pathways modulated by knock down of p53.
The Cignal Finder Cancer 10-Pathway Reporter Array showed that the knock
down of p53 gene expression down-regulates p53 signaling, while
up-regulating Notch, hypoxia and MAPK/ERK signaling in HEK-293H cells.
Interestingly, Notch signaling is known to be frequently deregulated in
human malignancies. Upregulation of Notch signaling by p53 RNA interference
suggests that Notch may function as a proto-oncogene.
HEK-293H cells were co-transfected with either p53 siRNA or a negative
control siRNA, in combination with each reporter assay and the negative
control from the Cancer 10-Pathway Reporter Array plate. Sixteen hours after
carrying out the reverse transfection, medium was changed to complete medium
(DMEM containing 10% of fetal bovine serum, 1% NEAA, 100 U/ml Penicillin and
100 µg/ml Streptomycin). After 48 hours of transfection, the dual-luciferase
assay was performed and results are expressed as fold change. The fold
change was calculated by dividing the normalized luciferase activities of
each pathway-focused reporter co-transfected with p53 siRNA by the
normalized luciferase activity of each pathway-focused reporter co-transfected
with the negative control siRNA. 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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